Compositions and methods of treating dry eye syndrome and other traumatized non-keratinized epithelial surfaces

ABSTRACT

The present disclosure relates to a composition including non-autologous plasma or serum and a polymer; method of treating ophthalmic diseases (e.g., dry eye syndrome) or moisturizing and/or repairing non-keratinized surfaces including nonhealing wounds with a composition including non-autologous plasma or serum and a polymer; and a therapeutic agent delivery device for delivery of the composition to the eye or a non-keratinized surface of a subject.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. Pat. Application No.17/454,117, filed on Nov. 9, 2021, which is a continuation of U.S. Pat.Application No. 16/309,810, filed Dec. 13, 2018, now U.S. Pat. No. US11,191,781 issued Dec. 7, 2021, which is a national stage entry, filedunder 35 U.S.C. § 371, of International Application No.PCT/US2017/037954, filed on Jun. 16, 2017, which claims the benefit ofpriority under 35 U.S.C. §119(e) to U.S. Provisional Pat. ApplicationNo. 62/350,772, filed Jun. 16, 2016, which contents are herebyincorporated by reference in their entireties.

BACKGROUND OF THE DISCLOSURE

Non-keratinized epithelial surfaces are found throughout much of thebody, except the limbs, and with the exception of the eye-ball, arepredominantly out of sight. Lubrication is continually bathingnon-keratinized epithelial surfaces that can range from a thin wateryaqueous solution to a thick, viscous fluid called mucus. Lubrication issecreted by exocrine glands that also range from unicellular glandsknown as goblet cells that secrete via an apocrine method, budding offtheir secretions in membrane-bound vesicles, to complex multicellularglands that discharge secretions via a duct. Non-keratinized epitheliallubrication contains glycoproteins, proteoglycans, peptides, and enzymesthat both promote and protect the health of epithelial cells. Not onlydoes the lubrication protect the non-keratinized epithelial frommechanical irritation from frictional trauma, it also providesantimicrobial properties, preventing invasion from infectious organisms,a constant threat to these exposed areas. Non-keratinized epithelialsurfaces form the lining of the cornea, conjunctiva, mouth, pharynx,esophagus, vocal cords, vagina, and cervix and play a vital role in thegastrointestinal, respiratory, and urogenital tracts.

These tissues are all sites of communication, where material andinformation are passed between the body and its environment. Because oftheir physiological functions of sensory activity (eyes, nose, mouth,and throat), gas exchange (lungs), food absorption (gut), andreproduction (uterus, vagina, and breast), the non-keratinizedepithelial surfaces are by necessity dynamic, thin, permeable barriersto the interior of the body. These properties make these tissuesparticularly vulnerable to subversion and breach by pathogens. Thisfragility, combined with the vital functions of non-keratinizedepithelial surfaces, has driven the evolution of specialized mechanismsfor their defense.

Dry eye syndrome is a large and growing problem and is very common withan estimated 25% of patients in general ophthalmology or optometryclinics reporting symptoms of dry eye syndrome. It has multiple causesthat result in an unstable tear film and rapid tear film breakup time.Dry eye syndrome is a multifactorial disease of the tears and ocularsurface that results in symptoms of blurry vision and discomfortincluding foreign body sensation, irritation, burning, and lightsensitivity. Dry eye and tear film instability can damage the ocularsurface. It is accompanied by increased osmolarity of the tear film andinflammation of the ocular surface.

Currently, there is no cure for dry eye syndrome and common treatmentsare targeted to merely manage the symptoms. The mainstay of conventionaltherapy is the application of artificial tears that increase moisture onthe ocular surface, and provide additional lubrication. A variety ofartificial tear formulations differ from each other in their electrolytecomposition, osmolarity, viscosity, presence or absence ofpreservatives, and compatible solutes. However, frequent application ofartificial tears containing preservatives to prevent contamination hasbeen found to include both toxic and allergic reactions, especially inpatients with sensitive eyes.

There is currently a need for an ophthalmic composition for thetreatment of dry eye syndrome. Among many challenges in developing dryeye therapy is the inability to deliver a composition that is retainedon the eye surface for a prolonged period. Topical administration,mostly in the form of eye drops, is usually employed to treat eyeanterior segment diseases. For most of the topically applied drugs, thesite of action is usually different layers of the cornea, conjunctiva,sclera, and the other tissues of the anterior segment such as the irisand ciliary body (anterior uvea). Upon administration, precornealfactors and anatomical barriers negatively affect the bioavailability oftopical formulations. Precorneal factors include solution drainage,blinking, tear film, tear turn over, and induced lacrimation. Tear filmdisplays a rapid restoration time of 2-3 min, and most of the topicallyadministered solutions are washed away within just 15-30 s afterinstillation. Considering all the precorneal factors, contact time withthe absorptive membranes is lower, which is considered to be the primaryreason for less than 5% of the applied dose reaching the intraoculartissues. Precorneal factors, however, are not a major obstacle intreating dry eye syndrome.

Various layers of cornea, conjunctiva, and sclera also play an importantrole in drug permeation. The cornea, the anterior most layer of the eye,is a mechanical barrier which limits the entry of exogenous substancesinto the eye and protects the ocular tissues. It can be mainly dividedinto the epithelium, stroma, and endothelium. Each layer offers adifferent polarity and a potential rate-limiting structure for drugpermeation. The corneal epithelium is lipoidal in nature which contains90% of the total cells in the cornea and poses a significant resistancefor permeation of topically administered hydrophilic drugs. Furthermore,superficial corneal epithelial cells are joined to one another bydesmosomes and are surrounded by ribbon-like tight junctional complexes(zonula occludens). Presence of these tight junctional complexes retardsparacellular drug permeation from the tear film into intercellularspaces of the epithelium as well as inner layers of the cornea.

The stroma, which comprises 90% of the corneal thickness, is made up ofan extracellular matrix and consists of a lamellar arrangement ofcollagen fibrils. The highly dense structure of the stroma poses asignificant barrier to permeation of lipophilic drug molecules.Endothelium is the innermost monolayer of hexagonal-shaped cells. Eventhough endothelium is a separating barrier between the stroma andaqueous humor, it helps maintain corneal transparency due to itsselective carrier-mediated transport and secretory function.Furthermore, the corneal endothelial junctions are leaky and facilitatethe passage of macromolecules between the aqueous humor and stroma.Thus, corneal layers, particularly the epithelium and stroma, areconsidered as major barriers for ocular drug delivery. The permeantshould have an amphipathic nature in order to permeate through theselayers.

Compared to the cornea, conjunctival drug absorption is considered to benonproductive due to the presence of conjunctival blood capillaries andlymphatics, which can cause significant drug loss into the systemiccirculation thereby lowering ocular bioavailability. Conjunctivalepithelial tight junctions can further retard passive movement ofhydrophilic molecules. The sclera, which is continuous with the corneaoriginates from the limbus and extends posteriorly throughout theremainder of the globe. The sclera mainly consists of collagen fibersand proteoglycans embedded in an extracellular matrix. Permeabilitythrough the sclera is considered to be comparable to that of the cornealstroma. The permeability of drug molecules across the sclera isinversely proportional to the molecular radius. Dextrans with linearstructures were less permeable as compared to globular proteins.Furthermore, the charge of the drug molecule also affects itspermeability across the sclera. Positively charged molecules exhibitpoor permeability presumably due to their binding to the negativelycharged proteoglycan matrix.

For treating dry eye syndrome a composition is needed that would havemaximal contact time with the absorptive membrane and reducedpermeability across the various layers of the cornea, conjunctiva, andsclera. Described herein are compositions for effective treatment of dryeye syndrome and ameliorating its symptoms.

SUMMARY OF THE DISCLOSURE

The current disclosure provides, inter alia, a freeze-dried compositionincluding non-autologous plasma or serum and a polymer; a method oftreating ophthalmic diseases (e.g., dry eye syndrome), the methodincluding applying the re-constituted freeze-dried composition; and anophthalmic therapeutic agent delivery device (100) including twochambers, wherein the first chamber (101) configured for storing afreeze-dried ophthalmic composition including of non-autologous plasmaor serum, and a and a second chamber (102) including a reconstitutionfluid. In one aspect the present disclosure provides compositions formoisturizing and/or repairing non-keratinized epithelial surfaces in oron the body of a subject. Examples include treatment of dry mouthsyndrome, vaginal dryness, diabetic ulcers and other chronic wounds. Inembodiments, a freeze-dried composition for treating dry eye syndrome orfor moisturizing and/or repairing non-keratinized epithelial surfaces isdisclosed, where the composition includes non-autologous plasma orserum, and a polymer having a molecular weight of 100 to 1200 kDA, orderivatives thereof. In embodiments the non-autologous plasma or serumof the freeze-dried composition may be of umbilical cord plasma orserum.

In embodiments, the disclosure provides a freeze-dried compositioncomprising non-autologous plasma or serum and a polymer that is abiopolymer, preferably a polysaccharide having a molecular weight of 100to 1200 kDa, preferably from 100 to 400 kDa, or a polysaccharide havinga molecular weight of 100, 250, or 400 kDa, wherein the polysaccharideis chitosan, or a hydrophobically modified chitosan. In embodiments, thefreeze-dried composition comprises serum or plasma in an amount asdescribed infra. In embodiments, the composition is biodegradable andbiocompatible. In embodiments, the freeze-dried composition is in theform of a cohesive solid state material, such as a disc, wafer, lens,pessary, wound dressing, or denture. In embodiments, the freeze-driedcomposition in the form of a solid material is further combined with asufficient amount of an aqueous liquid (e.g., a reconstitution fluid).In embodiments, liquid serum or plasma may be used as the reconstitutionfluid, either alone or diluted with an appropriate aqueous carrier, forexample water or saline. In embodiments, the aqueous liquid is anysolvent described herein. In exemplary embodiments, water or a salinesolution may be used, any additives may be included (e.g., glycine orascorbic acid). In embodiments, the disclosure provides a freeze-driedcomposition in the form of a cohesive solid state material along with areconstitution fluid which when mixed with the cohesive solid statematerial forms a cohesive sponge-like material. In another embodiment,the freeze-dried composition is in the form of a dry powder which can beformulated into a suitable dosage form for topical application to anon-keratinized epithelial surface, such as the surface of the eye,mouth, or vagina, or the surface of an external wound. Such powdercompositions may be formulated, for example, as a liquid, gel, orointment. Preferably, the compositions are sterile.

The freeze-dried plasma or serum and polysaccharide based compositionsdescribed here have numerous advantageous properties. For example, thecompositions are biodegradable and biocompatible with animal tissues,especially with non-keratinized epithelial surfaces, such as the surfaceof the eye, mouth, or vagina, or the surface of an external wound. Inembodiments, the compositions contain biologically active plasma orserum derived proteins whose dissolution from the site of application isslowed by the matrix-like structure of the composition. Accordingly, thedisclosure provides methods of promoting the healing of a damaged ortraumatized non-keratinized epithelial surface, including the surface ofan external wound, by applying an amount of a composition described hereto the surface.

In embodiments, the composition forms a matrix-like structure thatenables slow release of the biologically active components present inthe plasma or serum after application to a non-keratinized epithelialsurface, such as the eye for the treatment of ocular diseases anddisorders including, but not limited to, dry eye syndrome. In otheraspects, the freeze-dried composition described herein also promoteshealth of the underlying corneal epithelium, as evidenced for example bythe uptake of Rose Bengal dye according to standard assays for thedetection of corneal epithelial damage, e.g., in dry eye patients. Inembodiments, the compositions described here also improve tear break uptime, as measured by standard assays.

In embodiments the polymer of the freeze-dried composition may be alinear biopolymer. In some aspects, the polymer may be modified to besoluble in an aqueous solution. In embodiments, the polymer of thefreeze-dried composition may be a basic biopolymer with pH more than7.0. In some embodiments, the polymer may be cationic, anionic orzwitterionic and may be selected from chitosans, alginates, gelatins,and a combination thereof. In embodiments, the chitosan may be selectedfrom hydrophobically-modified chitosan, carboxymethyl chitosan, succinylchitosan, glycol chitosan, thiolated chitosan and a combination thereof.

In embodiments, the freeze-dried composition may further include anadditive. In embodiments, the additive may be selected from chitosan,alginate, gelatin, hyaluronic acid, gellan gum, dextran, polyethyleneglycol, polyethylene oxide, glucose, glucosamine, sodium chloride,polylactic acid, polylactic-co-glycolic acid and glycerol.

In embodiments, the non-autologous plasma or serum in the freeze-driedophthalmic composition may be at a concentration of about 0.005 to 100%by weight/vol. (w/v) of the composition.

In embodiments, the composition is reconstituted with a reconstitutionfluid. In embodiments, the reconstitution fluid may further include anadditive, wherein the additive is selected from ascorbic acid, glycine,and combination thereof. The reconstituted composition may have a finalpH of about 7.4.

In embodiments, the composition comprises non-autologous serum, and apolymer having a molecular weight of 100 to 1200 kDa. Alternatively, thecomposition is a freeze-dried composition comprising plasma or serum anda polysaccharide having a molecular weight of from 100 to 400 kDa. Inembodiments, the polysaccharide is selected from the group consisting ofchitosan, cellulose, dextrin, pectin, alginic acid, agar, agarose,carragenas, and derivatives thereof. In other embodiments, thepolysaccharide is chitosan, or a hydrophobically modified chitosan.

In embodiments, the freeze-dried composition described here is in theform of a cohesive solid state material having a defined shape. Inembodiments, the solid state material is sufficiently cohesive that itdoes not crack under low or moderate pressure. In embodiments, thefreeze-dried composition in the form of a cohesive solid state materialhaving a defined shape comprises serum or plasma and chitosan,preferably a hydrophobically modified chitosan as described infra. Inembodiments, the chitosan is hydrophobically modified with palmiticanhydride, as described infra. In embodiments, the cohesive solid statematerial is in the form of a disc, wafer, lens, pessary, wound dressing,or denture. In embodiments where a more cohesive material is desired,the freeze-dried composition comprises serum and chitosan, or ahydrophobically modified chitosan as described infra.

In embodiments, the freeze-dried composition described herein is in theform of a dry powder. In embodiments, the freeze-dried composition inthe form of a dry powder comprises serum or plasma and chitosan,preferably a hydrophobically modified chitosan as described infra. Inembodiments, the chitosan is hydrophobically modified with palmiticanhydride, as described infra. In embodiments, the freeze-driedcomposition comprises serum and chitosan, or a hydrophobically modifiedchitosan as described infra.

In accordance with any of the solid-state embodiments of thefreeze-dried compositions described here, the amount of serum or plasmacomponents in the dry composition is from about 50-90 weight percent(wt%) based on the total dry weight of the composition, and the amountof polysaccharide is from about 10-40 wt%. In embodiments, thefreeze-dried composition contains from about 60-80 wt% of serum orplasma components and from about 20-40 wt% polysaccharide, or from about70-80 wt% of serum or plasma components and from about 20-30 wt%polysaccharide.

In embodiments, the disclosure provides a freeze-dried compositioncomprising plasma or serum and a polysaccharide selected from chitosan,a hydrophobically modified chitosan, sucrose, or trehalose, wherein thecomposition comprises from about 60-90 wt% of the plasma or serumcomponents and from about 10-40 wt% of the polysaccharide.

In accordance with any of the solid-state embodiments of thefreeze-dried compositions described above, the composition may beformulated as a pharmaceutical composition, which may take the form of aliquid, suspension, ointment, or gel, depending on the carriers and, oneor more optional excipients, included in the formulation. Accordingly,the disclosure provides pharmaceutical compositions comprising thefreeze-dried compositions described here, which further comprise acarrier and one or more optional excipients. In embodiments, the carrieris an aqueous solution, for example water or buffered saline. Inembodiments, the one or more optional excipients may include, forexample, ethanol, a polyol, a surfactant, or a carbohydrate. Inembodiments, the one or more optional excipients may be selected fromsucrose and trehalose. In embodiments, the pharmaceutical composition issterile.

Provided herein is a method for treating dry eye syndrome or a method ofmoisturizing and/or repairing non-keratinized surfaces (e.g., dry mouthsyndrome, vaginal dryness, diabetic ulcers and other chronic wounds), ina subject in need thereof including: administering a compositionincluding non-autologous plasma or serum, and a polymer having amolecular weight of 100 to 1200 kDA, or derivatives thereof, and thenon-autologous plasma or serum is present in the composition an amounteffective to treat dry eye syndrome in the eye or moisturizing and/orrepairing non-keratinized surfaces (e.g., dry mouth syndrome, vaginaldryness, diabetic ulcers and other chronic wounds), respectively, of thesubject. In embodiments, the non-autologous plasma or serum has beenlyophilized and reconstituted.

Also provided herein are methods for treating a non-keratinizedepithelial surface of a subject in need of such treatment, the methodcomprising applying to the surface a composition comprisingnon-autologous plasma or serum, and a polymer having a molecular weightof 100 to 1200 kDA, or derivatives thereof, wherein the non-autologousplasma or serum is present in the composition in an amount effective totreat the non-keratinized epithelial surface of the subject. Inembodiments, the non-keratinized epithelial surface is selected from anocular surface, an oral surface, a vaginal surface, and the surface ofan external wound.

In embodiments the plasma or serum is of umbilical cord plasma or serum.In embodiments, the polymer is a linear biopolymer. In embodiments, thepolymer is modified to be soluble in an aqueous solution. Inembodiments, the polymer is a basic biopolymer with pH more than 7.0. Inembodiments, the polymer is polycationic or polyanionic.

In embodiments, the polymer is a chitosan, alginate, gelatin, or anycombination(s) thereof. In embodiments, the polymer is a chitosan. Inembodiments, the chitosan is selected from carboxymethyl chitosan,succinyl chitosan, glycol chitosan, thiolated chitosan and a combinationthereof. In embodiments, the chitosan is a hydrophobically modifiedchitosan. In embodiments, the hydrophobically modified chitosan isselected from chitosan lactate, chitosan salicylate, chitosanpyrrolidone carboxylate, chitosan itaconate, chitosan niacinate,chitosan formate, chitosan acetate, chitosan gallate, chitosanglutamate, chitosan maleate, chitosan aspartate, and chitosan glycolate.In embodiments, the hydrophobic substituents of the hydrophobicallymodified chitosan are provided by palmitic anhydride. In accordance withthese embodiments, the amount of the polymer in the freeze-driedcomposition is from about 10-50 wt%, or from about 20-40 wt%, or fromabout 20-30 wt%, based on the total dry weight of the composition. Inaccordance with these embodiments, the serum or plasma components willmake up the bulk of the remainder, or the entirety of the remainder ofthe dry weight. In embodiments, the amount of serum or plasma componentsin the composition is from about 50-90 wt%, or from about 60-80 wt%, orfrom about 70-80 wt%.

In embodiments, the reconstituted composition is formulated as asolution, suspension, semi-liquid, semi-solid gel, gel, emulsion,ointment, or cream. In embodiments, the reconstituted composition isadministered topically. In other embodiments, the reconstitutedcomposition is administered in the form of eye drops. In otherembodiments, the reconstituted composition may be administered orallyfor dry mouth syndrome. In embodiments, the oral administration for drymouth syndrome may be via a mouthwash, a mouth rinse, an oral rinse, amouth bath, and the like. In embodiments, administration to the cervicaland/or vaginal mucosa may be via a solution, gel, suspension, cream,ointment, foam, pessary, or tablet.

In other embodiments, the reconstituted composition may be administeredto the cervical and/or vaginal mucosa of a subject. In embodiments, thecervical and/or vaginal mucosa administration may be via a solution,gel, suspension, cream, ointment, foam, pessary, or tablet.Alternatively, the reconstituted composition may be administered bycontinuous release from a vaginal ring and the like.

In embodiments, the reconstituted composition may be administered todiabetic ulcers and other chronic wounds of a subject. In embodiments,administration to diabetic ulcers and other chronic wounds may beadministered topically or orally. In embodiments, topical administrationmay include a solution, suspension, semi-liquid, semi-solid gel, gel,emulsion, ointment, or cream. In embodiments, oral administration mayinclude a tablet.

In embodiments, the non-autologous plasma or serum in the composition isat a concentration of about 0.005 to 100% (w/v) of the composition. Inembodiments the non-autologous plasma or serum in the composition is ata concentration of about 75% (w/v) to 90% (w/v) and 10-30%. Inembodiments, the non-autologous plasma or serum in the composition is ata concentration of about 40% to 60% (w/v) of the composition.

In embodiments, the disclosure provides a freeze-dried compositioncomprising plasma or serum and a polysaccharide, preferably chitosan ora hydrophobically modified chitosan, for use in methods for treating anon-keratinized epithelial surface, including an external wound, of asubject in need of such treatment. In embodiments, the polysaccharide isa chitosan having a molecular weight of from 100 to 400 kDa. Inembodiments, the polysaccharide is a hydrophobically modified chitosanin which the hydrophobic substituent is provided by palmitic anhydride.

In embodiments, compositions including any of the compositions describedfor use in a subject in need of such treatment. In embodiments, thedisclosure provides a freeze-dried composition comprising plasma orserum and a polysaccharide, preferably chitosan or a hydrophobicallymodified chitosan, for use in methods for treating dry eye syndrome in asubject in need of such treatment. In embodiments, the polysaccharide isa chitosan having a molecular weight of from 100 to 400 kDa. Inembodiments, the polysaccharide is a hydrophobically modified chitosanin which the hydrophobic substituent is provided by palmitic anhydride.

Also provided herein is a therapeutic agent delivery device (100)including two chambers (101, 102), where the first chamber (101) isconfigured for storing a freeze-dried composition including ofnon-autologous plasma or serum, and a polymer having a molecular weightof 100 to 1200 kDA, or derivatives thereof, and a second chamber (102)is configured for storing a reconstitution fluid.

In embodiments, the non-autologous plasma or serum is of umbilical cordplasma or serum. In embodiments, the polymer is a linear biopolymer. Inembodiments, the polymer is modified to be soluble in an aqueoussolution. In embodiments, the polymer is a basic biopolymer with pH morethan 7.0. In embodiments the polymer is polycationic. In embodiments,the polymer is selected from the chitosans, alginates, gelatins, and acombination thereof. In embodiments, the linear biopolymer is achitosan. In embodiments, the chitosan is selected from carboxymethylchitosan, succinyl chitosan, glycol chitosan, thiolated chitosan and acombination thereof. In embodiments, freeze-dried ophthalmic compositionis reconstituted with the reconstitution fluid such that the final pH isabout 7.4.

In embodiments the two chambers (101, 102) of the therapeutic agentdelivery device (100) are disrupted electromechanically or mechanically,thereby allowing mixing of the freeze-dried composition and thereconstitution fluid, thereby generating a reconstituted serum or plasmaformulation.

In embodiments, the therapeutic agent delivery device (100) of thepresent disclosure includes a freeze-dried composition in the firstchamber (101) and a reconstitution fluid in the second chamber (102),and upon reconstitution, the reconstituted composition is delivered to asubject.

In embodiments, the therapeutic agent delivery device (100) of thepresent disclosure includes the reconstituted composition which is fortreating dry eye syndrome, or moisturizing and/or repairing anon-keratinized surface.

Other features and advantages of the disclosure will be apparent fromthe following detailed description and claims.

Unless noted to the contrary, all publications, references, patentsand/or patent applications reference herein are hereby incorporated byreference in their entirety for all purposes.

BRIEF DESCRIPTION OF THE DRAWINGS

The patent or application file contains at least one drawing executed incolor. Copies of this patent or patent application publication withcolor drawing(s) will be provided by the Office upon request and paymentof the necessary fee.

FIG. 1 shows a schematic of a therapeutic agent delivery device (100)

FIG. 2A shows a Differential Scanning Calorimetry (DSC) thermogram of a5% sucrose formulation with a serum sample

FIG. 2B shows a Differential Scanning Calorimetry (DSC) thermogram of a5% sucrose formulation with a plasma sample

DETAILED DESCRIPTION OF THE DISCLOSURE

Provided herein, inter alia, is a freeze-dried ophthalmic compositionincluding non-autologous plasma or serum and a polymer; a method oftreating ophthalmic diseases (e.g., dry eye syndrome) of a subject inneed thereof, the method including applying the reconstitutedfreeze-dried ophthalmic composition (e.g., non-autologous plasma orserum and a polymer); and a delivery device (100) including thefreeze-dried ophthalmic composition and a reconstitution fluid.

Definitions

The following definitions are included for the purpose of understandingthe present subject matter and for constructing the appended patentclaims. Abbreviations used herein have their conventional meaning withinthe chemical and biological arts.

Unless defined otherwise, all technical and scientific terms used hereinhave the meaning commonly understood by a person skilled in the art towhich this disclosure belongs. The following references provide one ofskill with a general definition of many of the terms used in thisdisclosure: The Cambridge Dictionary of Science and Technology (Walkered., 1988); The Glossary of Genetics, 5th Ed., R. Rieger et. al. (eds.),Springer Verlag (1991); and Hale & Marham, The Harper Collins Dictionaryof Biology (1991). As used herein, the following terms have the meaningsascribed to them below, unless specified otherwise.

Unless specifically stated or obvious from context, as used herein, theterm “or” is understood to be inclusive. Unless specifically stated orobvious from context, as used herein, the terms “a”, “an”, and “the” areunderstood to be singular or plural.

Unless specifically stated or obvious from context, as used herein, theterm “about” is understood as within a range of normal tolerance in theart, for example within 2 standard deviations of the mean. About can beunderstood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%,0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise clear fromcontext, all numerical values provided herein are modified by the termabout. About with respect to concentration range of the compositions ofthe current disclosure also refers to any variation of a stated amountor range which would be an effective amount or range.

As used herein, “additive” can include any additional components thatmay be added to the composition as described herein. The additive may beadded prior to freeze-drying or prior to reconstitution. One or moreadditives may be added to the composition. The additive may includecomponents that have been classified by the FDA, and in embodiments maybe classified as Ophthalmic Drug Products for Over-The-Counter Human Use[Title 21, Volume 5]. Exemplary additives may include astringents,buffering agents, demulcents, emollients, eyewash, eye lotion,irrigating solutions, hypertonic agents, isotonic agents andvasoconstrictors. Additives in the current disclosure may be used in anysuitable amount.

As used herein, the term “administering” means oral administration,administration as a suppository, topical contact (e.g., eye drops or aspray), intravenous, parenteral, intraperitoneal, intramuscular,intralesional, intrathecal, intranasal or subcutaneous administration,or the implantation of a slow-release device, e.g., a mini-osmotic pump,to a subject. Administration is by any route, including parenteral andtransmucosal (e.g., buccal, sublingual, palatal, gingival, nasal,vaginal, cervical, rectal, or transdermal). Parenteral administrationincludes, e.g., intravenous, intramuscular, intra-arteriole,intradermal, subcutaneous, intraperitoneal, and intraventricular.

Administering may further include via oral administration (e.g., for thetreatment of dry mouth syndrome). In some examples, administrationorally may include a mouthwash, a mouth rinse, an oral rinse, a mouthbath, and the like. Cervical and/or vaginal mucosa administration may bevia a solution, a gel, a suspension, a cream, an ointment, a foam, apessary, or a tablet. In one aspect, the reconstituted composition maybe administered to the cervical and/or vaginal mucosa of a subject. Thecervical and/or vaginal mucosa administration may be via a solution,gel, suspension, cream, ointment, foam, pessary, or tablet.Alternatively, the reconstituted composition may be administered bycontinuous release from a vaginal ring and the like. Administration mayinclude administration to diabetic ulcers and other chronic wounds of asubject. Administration to diabetic ulcers and other chronic wounds maybe administered topically or orally. Topical administration may includea solution, suspension, semi-liquid, semi-solid gel, gel, emulsion,ointment, or cream. Oral administration may include a tablet.

As used herein, the term “autologous” refers denoting, relating to, orinvolving tissues or cells that are from one’s own sample (i.e., apatient’s own blood or blood components). Autologous plasma or serum isusually not readily available from a blood bank or a central repositoryunless it has been previously donated by the recipient.

“Biopolymers” as used herein may refer to polymers that are produced byliving organisms; they are “polymeric biomolecules.” Since they arepolymers, biopolymers contain repeat monomeric units that are covalentlybonded to form larger structures (i.e., linear biopolymers). Exemplarybiopolymers may include cellulose, starch, lignin, chitin, and variouspolysaccharides. These materials and their derivatives offer a widerange of properties and applications. Natural polymers tend to bereadily biodegradable, although the rate of degradation is generallyinversely proportional to the extent of chemical modification.

“Chitosan” as used herein refers to the principle derivative of chitin,is formed from chitin by deacetylation in the presence of alkali.Chitin, a polymer of N-acetylglucosamine, is a cellulose-like biopolymerthat is the main component of crustaceans (e.g. shrimp, crab, lobster)and is also present in the exoskeleton and the cell wall of fungi,insects and yeast. Chitosan is a linear polysaccharide composed of (1 →4)-2-acetamido-2-deoxy-b-D-glucan (N-acetyl D-glucosamine) and (1 →4)-2-amino-2-deoxyb-D-glucan (D-glucosamine) units. Chitosan hasnumerous biological properties, including antimicrobial activity,hemostatic activity, and acceleration of wound healing,tissue-engineering scaffolds, drug delivery, and antitumor activity.Chitosan’s inherent anti-microbial properties can be a crucial asset indecreasing the chance of infection and/or contamination. Additionally,chitosan, when biological burden is removed, is biodegradable andbiocompatible with low toxicity to mammalian cells.

Chitosan has positive charges along its backbone that cause it tointeract electrostatically with negatively charged pharmaceuticals(e.g., blood cells), thus creating a sticky interface between chitosanand any potential ocular wound providing hemostasis. Carboxymethylchitosan, succinyl chitosan, glycol chitosan, thiolated chitosan can beused to reduce the requirements for acid solubilization. Chitosan is alinear polymer of glucosamine units. Structurally, it is distinguishedfrom cellulose by the presence of the primary amine group. Chitosan isavailable commercially in various grades and average molecular weights(e.g., Sigma Aldrich®).

Chitosan may be derivatized by utilizing the reactivity on the aminogroup and or hydroxyl groups. Some exemplary chitosan derivatives mayinclude N-pathaloylation of chitosan, dendronized chitosan-sialic acidhybrids, methylthiocarbamoyl and phenylthiocarbamoyl chitosans,lactic/glycolic acid chitosan hydrogels, or nanocomposite from naturalpolysaccharide chitosan. In some aspects, chitosan may behydrophobically-modified. A chitosan can be hydrophobically-modifiedusing standard techniques in the art. In certain aspects, chitosans canbe hydrophobically-modified by reaction of alkyl (or aryl) aldehydeswith primary amine groups along the chitosan backbone (i.e., in a 50/50(v/v)% of aqueous acetic acid and ethanol). After reaction, theresulting Schiff bases or imine groups can be reduced to stablesecondary amines by dropwise addition of a reducing agent. In otheraspects, the chitosan can be hydrophobically-modified through theaddition of palmitic anhydride to the chitosan.

By “co-administer” it is meant that a composition described herein isadministered at the same time, just prior to, or just after theadministration of additional therapies. The composition of thedisclosure can be administered alone or can be co-administered with asecond composition/therapeutic agent to a subject. Co-administration ismeant to include simultaneous or sequential administration of thecomposition individually or in combination with a secondcomposition/therapeutic agent.

In this disclosure, “comprises,” “comprising,” “containing” and “having”and the like can have the meaning ascribed to them in U.S. Patent lawand can mean “includes,” “including,” and the like; “consistingessentially of” or “consists essentially” likewise has the meaningascribed in U.S. Patent law and the term is open-ended, allowing for thepresence of more than that which is recited so long as basic or novelcharacteristics of that which is recited is not changed by the presenceof more than that which is recited, but excludes prior art embodiments.

As used herein, “concurrent administration” includes overlapping induration at least in part. For example, when two agents (e.g., any ofthe compositions described herein) are administered concurrently, theiradministration occurs within a certain desired time. The compositions’administration may begin and end on the same day. The administration ofone composition can also precede the administration of a secondcomposition by day(s) as long as both compositions are taken on the sameday at least once. Similarly, the administration of one composition canextend beyond the administration of a second composition as long as bothcompositions are taken on the same day at least once. The compositionsdo not have to be taken at the same time each day to include concurrentadministration.

As used herein, the term, “cream” may refer to a thick (high viscosity)liquid or semi-liquid that may be used for therapeutic treatment of adisease, syndrome, or condition (i.e., dry eye syndrome). Examples ofsuitable creams include, but are not limited to, water-in-oil and oil-in-water emulsions. Water-in-oil creams may be formulated by using asuitable emulsifying agent with properties similar, but not limited, tothose of the fatty alcohols such as cetyl alcohol or cetostearyl alcoholand to emulsifying wax. Oil-in-water creams may be formulated using anemulsifying agent such as cetomacrogol emulsifying wax. Suitableproperties include the ability to modify the viscosity of the emulsionand both physical and chemical stability over a wide range of pH. Thewater soluble or miscible cream base may contain a preservative systemand may also be buffered to maintain an acceptable physiological pH.

As used herein, “digital mechanism” may refer to a means by which anon-permeable membrane can be broken on demand by a user, wherein theuser may press a button on the delivery device (100) and therebyallowing mixing of the Chamber A (101) and Chamber B (102) within thedelivery device (100).

The term, “dry eye” or “dry eye syndrome” may refer to an ophthalmicsyndrome or ocular surface condition. Dry eye or dry eye syndromeinclude keratoconjunctivitis sicca (KCS), dysfunctional tear syndrome,lacrimal keratoconjunctivitis, evaporative tear deficiency, aqueous teardeficiency, and LASIK-induced neurotrophic epitheliopathy (LNE). Dry eyeand tear film instability can damage the ocular surface. It isaccompanied by increased osmolarity of the tear film and inflammation ofthe ocular surface. An increase in tear osmolarity, which causes ocularsurface inflammation, is thought to be the central pathogenic mechanismof dry eye.

Dry eye syndrome is a large and growing problem and is very common withan estimated 25% of patients in general ophthalmology or optometryclinics reporting symptoms of dry eye. It has multiple causes thatresult in an unstable tear film and rapid tear film breakup time. It isknown that the incidence of dry eye syndrome increases with age and hasa higher prevalence in women relative to men. Currently, there is nocure for dry eye syndrome. Common treatments are targeted to manage thesymptoms. The mainstay of conventional therapy is the application ofartificial tears that increase moisture on the ocular surface, andprovide additional lubrication. A variety of artificial tearformulations differ from each other in their electrolyte composition,osmolarity, viscosity, presence or absence of preservatives, andcompatible solutes. However, frequent application of artificial tearscontaining preservatives to prevent contamination has been found toinclude both toxic and allergic reactions, especially in patients withsensitive eyes. Additional therapies for dry eye syndrome are alsoinadequate. These therapies include immunomodulators (e.g. Restasis®,Xiidra®) and corticosteroids. They tend to only address the symptoms,rather than the cause of the disease with corticosteroids in particularhaving the risk of causing cataracts and glaucoma.

As used herein, an “effective amount” or “therapeutically effectiveamount” is that amount sufficient to affect a desired biological effect,such as beneficial results, including clinical results. As such, an“effective amount” depends upon the context in which it is beingapplied. An effective amount may vary according to factors known in theart, such as the disease state, age, sex, and weight of the individualbeing treated. Several divided doses may be administered daily or thedose may be proportionally reduced as indicated by the exigencies of thetherapeutic situation. In addition, the compositions/formulations ofthis disclosure can be administered as frequently as necessary toachieve a therapeutic amount.

As used herein, “electrochemical mechanism” may refer to a means bywhich a non-permeable membrane can be broken on demand by a user,wherein the user introduces an electromechanical signal (i.e. voltage orcurrent signals) on the delivery device (100) and thereby allowingmixing of the Chamber A (101) and Chamber B (102) within the deliverydevice (100).

“Emulsion” as used herein may include a fine dispersion of minutedroplets of one liquid in another in which it is not soluble ormiscible. Alternatively, “emulsion” may refer to a mixture of two ormore liquids that are normally immiscible (unmixable or unbendable).Emulsions are part of a more general class of two-phase systems ofmatter called colloids.

The term, “freeze-dried” or “lyophilized” refers to a dehydrationprocess typically used to preserve a perishable material or make thematerial more convenient for transport. Freeze-drying works by freezingthe material and then reducing the surrounding pressure to allow thefrozen water in the material to sublimate directly from the solid phaseto the gas phase. If a freeze-dried substance is sealed to prevent thereabsorption of moisture, the substance may be stored at roomtemperature without refrigeration, and be protected against spoilage formany years. Preservation is possible because the greatly reduced watercontent inhibits the action of microorganisms and enzymes that wouldnormally spoil or degrade the substance. Freeze-drying also causes lessdamage to the substance than other dehydration methods using highertemperatures. Freeze-drying does not usually cause shrinkage ortoughening of the material being dried. Freeze-dried products can berehydrated (reconstituted) much more quickly and easily because theprocess leaves microscopic pores. The pores are created by the icecrystals that sublimate, leaving gaps or pores in their place. This isespecially important when it comes to pharmaceutical uses. By removingthe water from the material and sealing the material in a vial, thematerial can be easily stored, shipped, and later reconstituted to itsoriginal form for injection. Freeze-drying can also be used to increasethe shelf life of some pharmaceuticals for many years.

As used herein, “gamma irradiation” refers to a means of pathogenreduction. Gamma rays are a form of electromagnetic radiation withhigher energy than x-rays. The primary industrial sources of gamma raysare radionuclide elements such as Cobalt 60, which emit gamma raysduring radioactive decay. Gamma rays pass readily through plastics andkill bacteria by breaking the covalent bonds of bacterial DNA. They aremeasured in units called kiloGrays (kGy). Gamma irradiation between, forexample, 10 and 50 kGy, may be performed to the serum or plasma at 4° C.before and/or after it is freeze-dried. This inactivates potentialpathogens including viruses that may not have been previouslyeradicated. An additive (i.e., ascorbic acid or glycine) may be added asa protectant for plasma proteins against gamma irradiation and for pHbalance. In some examples, gamma irradiation may be performed to theserum or plasma before it is freeze-dried. The gamma irradiated liquidserum or plasma may be stored and frozen at minus 70° C. for futurepathogen reduction and/or freezing drying. In other examples, gammairradiation may be performed to the serum or plasma after it isfreeze-dried. The gamma irradiated freeze-dried serum or plasma may bestored and frozen at minus 70° C. for future pathogen reduction.

The term, “gel” as used herein may refer to a material which is not areadily flowable liquid and not a solid, i.e., semi-solid. Gels may beformed from naturally occurring or synthetic materials. The gels can benon-ordered to slightly ordered showing some birefringence, liquidcrystal character. Gels maybe produced using a suitable gelling agentincluding, but not limited to, gelatin, tragacanth, or a cellulosederivative and may include glycerol as a humectant, emollient, andpreservative. Gels are administered topically or, for example, aftershaking, in the form of a hydrogel as an eye drop.

As used herein, “intermittent administration” includes theadministration of a composition for a period of time (which can beconsidered a “first period of administration”), followed by a timeduring which the composition is not taken or is taken at a lowermaintenance dose (which can be considered “off-period”) followed by aperiod during which the composition is administered again (which can beconsidered a “second period of administration”). Generally, during thesecond phase of administration, the dosage level of the composition willmatch that administered during the first period of administration butcan be increased or decreased as medically necessary.

“Jelly” according to the current disclosure is a class of gels, whichare semisolid systems that consist of suspensions made up either smallinorganic particles or large organic molecules interpenetrated by aliquid, in which the structural coherent matrix contains a high portionof liquid, usually water.

“Liquid” as used herein is a dosage form consisting of a composition inits liquid state. A liquid is pourable; it flows and conforms to itscontainer at room temperature. Liquids display Newtonian orpseudoplastic flow behavior. In embodiments, a “semi-liquid” as usedherein may have properties of both a liquid and another formulation(i.e., a suspension, an emulsion, a solution, a cream, a gel, a jelly,and the like).

The term “linear biopolymer” as used herein may refer to polymersproduced by living organisms (e.g., biopolymers) that are connected in astraight chain of repeat monomeric subunits.

“Lubricating” or “lubricant” as used herein may refer to reducing thefriction between two surfaces (i.e., non-keratinized surfaces).Lubricants prevent ingredients from sticking together. Common mineralslike talc or silica, and fats, e.g. vegetable stearin, magnesiumstearate or stearic acid are the most frequently used lubricants intablets or hard gelatin capsules. Lubricants are agents added in smallquantities to tablet and capsule formulations to improve certainprocessing characteristics. Other roles of lubricants include ananti-adherent role and a glidant role (i.e., to enhance product flow).Lubricants can be both hydrophilic and hydrophobic. In general, mostwidely used lubricants are hydrophobic. Hydrophobic lubricants aregenerally good lubricants and are usually effective at relatively lowconcentrations. Many also have both anti-adherent and glidantproperties. For these reasons, hydrophobic lubricants are used much morefrequently than hydrophilic compounds. Examples include magnesiumstearate.

The term “moisturizing”, as used herein, refers to improving hydrationof a non-keratinized surface, such that water-binding capacity of thesurface increases. The term “moisturize” or derivatives thereof, relatesto the conversion or enhancement of the water contents ofnon-keratinized surfaces of a subject. The term “humectant” is usedherein in its usual sense, refers to water-soluble, physiologicallyacceptable, substances which are hygroscopic and capable ofspontaneously absorbing water vapor. “Occlusion” is a term used hereinto indicate the trapping of water in the non-keratinized surfaces by alayer of water-impervious fatty substance. Exemplary vaginalmoisturizers include, but are not limited to, Replens®, K-Y Liquibeads®,Lubrin®, Astroglide Silken Secret® and Vitamin E gel. Examples of oralmoisturizes (i.e. for dry mouth) may include, but are not limited to,artificial saliva products, saliva stimulants, Salese Soothing®,Orajel®, and Eucerin® cream. Exemplary moisturizers for diabetic ulcersand/or chronic wounds may include any saline or similar dressings thatprovide a moist environment, TriDerma® MD Ulcer Healing Cream, orNeoteric Diabetic Healing Cream®.

The term “non-autologous” as used herein may refer to denoting, relatingto, or involving tissues or cells (including plasma or serum) that arenot originated from the recipient (i.e., a subject in need of atreatment with the tissues or cells). In contrast, “autologous,” as usedherein, is in its ordinary meaning cells, tissues (including plasma orserum) obtained from the same individual, i.e., the recipient’s ownplasma or serum, and does not include plasma or serum originating fromanother individual (i.e., umbilical cord plasma or serum may be includedas non-autologous). Autologous plasma or serum is usually not readilyavailable from a blood bank or a central repository unless it has beenpreviously donated by the recipient. The term “allogenic” or“homologous” is used here by its ordinary meaning, i.e., denoting,relating to, or involving tissues or cells (including plasma or serum)that are genetically dissimilar and hence immunologically incompatible,although from individuals of the same species. Allogenic plasma or serumis, therefore, encompassed by the term “non-autologous” plasma or serum.In some examples, non-autologous plasma or serum may be purchased from aBlood Bank.

As used herein, “ocular surface disorder” “ophthalmic disease,”“ophthalmic disorder,” and the like, includes, but is not limited to,dry eyes, epithelial defects, Superior limbic keratoconjunctivitis,keratoconjunctivitis sicca, Neurotrophic keratopathy, Sjögren’ssyndrome, Stevens-Johnson syndrome, Ocular cicatricial pemphigoid,Medicamentosa, Graft-versus-host disease, and corneal ulcerations anderosions.

The term, “non-permeable membrane” describes a barrier separating twoenvironments (i.e. Chamber A (101) and Chamber B (102) of the ophthalmictherapeutic agent delivery device (100) including the freeze-driednon-autologous plasma or serum and a reconstitution fluid,respectively), whereby nothing passes in-between the two environments.The non-permeable membrane as described herein may be broken by a user,for example, by a digital mechanism, an electromechanical mechanism, oragitation or twisting.

“Ocular surface” as used herein includes the cornea and the conjunctiva.The ocular surface is covered by a thin layer of fluid or tear film. Thetear film is not only responsible for the majority of the refractivepower of the eye (approximately two-thirds) and clear vision, it is alsoresponsible for nourishing the cells on the surface of the eye andpreventing infection. It is the first line of defense against foreignpathogens and supports healing. Since the surface epithelium of the eyeis similar to other external surfaces of the body and regularlyrefreshes itself by shedding its epithelial cells, the tears play avital role in the health of this process. The surface of the eye cansuffer many kinds of diseases. One of most common diseases of thesurface of the eye is dry eye syndrome.

“Ocular surface conditions” as used herein may include any condition ofthe surface of the eye (e.g., conjunctiva and cornea). The conditionsmay include, but are not limited to, dry eye, epithelial defects,Superior limbic keratoconjunctivitis, Keratoconjunctivitis sicca,Neurotrophic keratopathy, Sjögren’s syndrome, Stevens-Johnson syndrome,Ocular cicatricial pemphigoid, Medicamentosa and Graft-versus-hostdisease.

“Ocular surface surgery″ may include penetrating keratoplasty andrefractive surgery laser-assisted in situ keratomileusis (LASIK), laserepithelial keratomileusis (LASEK), or photorefractive keratectomy (PRK).

As used herein the term “ophthalmic composition” refers to a compositionintended for application to the eye or its related or surroundingtissues such as, for example, the eyelid or onto the cornea. The termalso includes compositions intended to therapeutically treat conditionsof the eye itself or the tissues surrounding the eye. The ophthalmiccomposition can be applied topically or by other techniques, known topersons skilled in the art, such as injection to the eye. Examples ofsuitable topical administration to the eye include administration in eyedrops and by spray formulations. A further suitable topicaladministration route is by subconjunctival injection. The compositionscan also be provided to the eye periocularly or retro-orbitally.

As used herein, an “ophthalmic therapeutic agent delivery device” mayincorporate the compositional elements described herein (e.g., thefreeze-dried non-autologous plasma or serum polymer, and reconstitutionfluid).

As used herein, the term “ointment” may refer to a highly viscous liquidor semi-liquid formulation that may be used for therapeutic treatment ofa disease, syndrome, or condition (i.e., dry eye syndrome).

“Patient,” “subject,” “patient in need thereof,” and “subject in needthereof” are herein used interchangeably and refer to a living organismsuffering from or prone to a disease or condition that can be treated byadministration using the methods and compositions provided herein. Nonlimiting examples include humans, other mammals, bovines, rats, mice,dogs, monkeys, goat, sheep, cows, deer, and other non-mammalian animals.In some embodiments, a patient is human.

“Pessaries” or “pessary” as used herein may refer to a solid unit-doseform suitably shaped for insertion into the vagina and may either becomposed of a base that melts at body temperature or which dissolveswhen in contact with mucous secretions. Examples of suitable basesinclude, but are not limited to, theobroma oil, synthetic fat bases(e.g. Witepsol®), polyethylene glycols (macrogols), and glycerolsuppository basis.

“Plasma” as used herein may refer to a fluid substance in blood that isthe pale straw colored liquid component of blood that normally holds theblood cells in whole blood in suspension; this in turn makes plasma theextracellular matrix of blood cells. It mostly consists of water (up to95% by volume), and contains dissolved proteins, (including, forexample, serum albumins, globulins, and fibrinogen), glucose, clottingfactors, electrolytes (e.g., Na⁺, Ca₂ ⁺, Mg₂ ⁺, HCO₃ ⁻, Cl⁻), hormones,and carbon dioxide (plasma being the main medium for excretory producttransportation). Plasma also serves as the protein reserve of the humanbody. It plays a vital role in an intravascular osmotic effect thatkeeps electrolytes in balanced form and protects the body from infectionand other blood disorders. In embodiment, “plasma” is an engineeredcomposition including components of natural human plasma.

The term “polymer” as referred to herein is meant as a macromolecule,composed of many repeated subunits called monomers. The word polymerdesignates an unspecified number of monomer units. When the number ofmonomers is very large, the compound is sometimes called a high polymer.Polymers are not restricted to monomers of the same chemical compositionor molecular weight and structure. Some natural polymers are composed ofone kind of monomer. Most natural and synthetic polymers, however, aremade up of two or more different types of monomers. Such polymers areknown as copolymers. Polymers can be linear or branched. For example, alinear polymer characterized by a repetition of ester groups along thebackbone chain is called a polyester.

Exemplary natural polymeric materials include shellac, amber, wool,silk, rubber, and cellulose. Non-natural (e.g., synthetic) polymersinclude synthetic rubber, phenol formaldehyde resin, neoprene, nylon,polyvinyl chloride, polystyrene, polyethylene, polypropylene, andsilicone.

Additional synthetic polymers that can be used include biodegradablepolymers such as poly(lactide) (PLA), poly(glycolic acid) (PGA),poly(lactide-co-glycolide) (PLGA), poly(caprolactone), polycarbonates,polyamides, polyanhydrides, polyamino acids, polyortho esters,polyacetals, polycyanoacrylates and degradable polyurethanes, andnon-erodible polymers such as polyacrylates, ethylene-vinyl acetatepolymers and other acyl substituted cellulose acetates and derivativesthereof, non-erodible polyurethanes, polystyrenes, polyvinyl chloride,polyvinyl fluoride, poly(vinyl imidazole), chlorosulphonatedpolyolifins, polyethylene oxide, polyvinyl alcohol, teflon®, and nylon.A non-absorbable polyvinyl alcohol sponge is available commercially asIvalon™, from Unipoint Industries.

The term, “polycation” or “cationic” refers to a compound or moleculedescribed herein (e.g., polymers such as chitosan and chitosanderivatives) that may be protonated and thereby having an overallpositive charge. The protonation may be, for example, on the aminogroups of the compound or molecule (e.g., polymers such as chitosan andchitosan derivatives) and can then subsequently form ionic complexeswith a wide variety of natural or synthetic anionic species, includingfor example, lipids, proteins, DNA and other negatively chargedsynthetic polymers (e.g., poly (acrylic acid)). Chitosan, for example,is the only positively charged, naturally occurring polysaccharide.

Polysaccharides are polymeric carbohydrate structures, formed fromrepeating units joined together with glycosidic bonds. Their structuresare often linear, but may contain various degrees of branching. Innature, polysaccharides have various resources from algal origin, plantorigin, microbial origin and animal origin. Polysaccharides have ageneral formula of C_(x)(H₂O)_(y) where x is usually a large numberbetween 200 and 2500. Considering that the repeating units in thepolymer backbone are often six-carbon monosaccharides, the generalformula can also be represented as (C₆H₁₀O₅)n where 40≤ n ≤3000.

Examples of monosaccharides are glucose, fructose, and glyceraldehyde.Examples of naturally occurring polysaccharides are: cellulose, dextrin,pectin, alginic acid, agar, agarose, and carragenas. Naturally occurringpolysaccharides are usually acidic in nature.

In embodiments, polysaccharides may contain more than ten monosaccharideunits. Polysaccharides are an important class of biological polymers.Their function in living organisms is usually either structure- orstorage-related. Cellulose and chitin are examples of structuralpolysaccharides. Cellulose is used in the cell walls of plants and otherorganisms, and is said to be the most abundant organic molecule onEarth. Chitin has a similar structure to cellulose, but hasnitrogen-containing side branches, increasing its strength. It is foundin arthropod exoskeletons and in the cell walls of some fungi. It alsohas multiple uses, including surgical threads. Polysaccharides alsoinclude callose or laminarin, chrysolaminarin, xylan, arabinoxylan,mannan, fucoidan and galactomannan.

The terms “prevent,” “preventing,” or “prevention,” “prophylactictreatment” and the like, refer to reducing the probability of developinga disorder or condition in a subject, who does not have, but is at riskof or susceptible to developing a disorder or condition. The preventionmay be complete (i.e., no detectable symptoms) or partial, so that fewersymptoms are observed than would likely occur absent treatment. Theterms further include a prophylactic benefit. For a disease or conditionto be prevented, the compositions may be administered to a patient atrisk of developing a particular disease, or to a patient reporting oneor more of the physiological symptoms of a disease, even though adiagnosis of this disease may not have been made.

“Psoralen” or “psoralene” is the parent compound in a family of naturalproducts known as furocoumarins. It is structurally related to coumarinby the addition of a fused furan ring, and may be considered as aderivative of umbelliferone. It is widely used in PUVA (psoralen + UVA)treatment for psoriasis, eczema, vitiligo, and cutaneous T-celllymphoma. Psoralens are often used for the inactivation of pathogens inblood products. The synthetic amino-psoralen, amotosalen HCl, has beendeveloped for the inactivation of infectious pathogens (bacteria,viruses, protozoa) in platelet and plasma blood components prepared fortransfusion support of patients. A photoactive solvent or detergent ismixed into the serum or plasma and exposed to ultraviolet light. Withamotosalen HCl, for example, photoactivation occurs when the mixture isilluminated with UVA treatment at 3 J/cm². This reduces the risk oftransfusion-associated transmission of viruses, bacteria, and parasitesthat theoretically may be present.

Ranges can be expressed herein as from “about” one particular value,and/or to “about” another particular value. When such a range isexpressed, another aspect includes from the one particular value and/orto the other particular value. Similarly, when values are expressed asapproximations, by use of the antecedent “about,” it is understood thatthe particular value forms another aspect. It is further understood thatthe endpoints of each of the ranges are significant both in relation tothe other endpoint, and independently of the other endpoint. It is alsounderstood that there are a number of values disclosed herein, and thateach value is also herein disclosed as “about” that particular value inaddition to the value itself. It is also understood that throughout theapplication, data are provided in a number of different formats and thatthis data represent endpoints and starting points and ranges for anycombination of the data points. For example, if a particular data point“10” and a particular data point “15” are disclosed, it is understoodthat greater than, greater than or equal to, less than, less than orequal to, and equal to 10 and 15 are considered disclosed as well asbetween 10 and 15. It is also understood that each unit between twoparticular units are also disclosed. For example, if 10 and 15 aredisclosed, then 11, 12, 13, and 14 are also disclosed.

Ranges provided herein are understood to be shorthand for all of thevalues within the range. For example, a range of 1 to 50 is understoodto include any number, combination of numbers, or sub-range from thegroup consisting 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34,35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 aswell as all intervening decimal values between the aforementionedintegers such as, for example, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8,and 1.9. With respect to sub-ranges, “nested sub-ranges” that extendfrom either end point of the range are specifically contemplated. Forexample, a nested sub-range of an exemplary range of 1 to 50 may include1 to 10, 1 to 20, 1 to 30, and 1 to 40 in one direction, or 50 to 40, 50to 30, 50 to 20, and 50 to 10 in the other direction.

“Reconstitution” as referred to herein means in its ordinary sense, toreturn a dehydrated or concentrated composition to the liquid state byadding a liquid.

As used herein, “reconstitution fluid” may refer to any solventdescribed herein used to reconstitute the freeze-dried composition ofthe current disclosure. In exemplary embodiments, water or a salinesolution (e.g. 0.9% w/v sodium chloride for injection) may be used, anyadditives may be included, into the reconstitution fluid (i.e., glycineor ascorbic acid, or chitosan, alginate, and gelatin), and serum mayalso be used as a reconstitution fluid. In embodiments, the solventsused for the reconstitution fluid may be sterile.

The reconstituted freeze-dried composition may be reconstituted as asolution, suspension, semi-liquid, emulsion, ointment, cream, orsemi-solid gel.

As used herein, “salts” or “salt form” or “pharmaceutically acceptedsalts” may include base addition salts (formed with free carboxyl orother anionic groups) which are derived from inorganic bases such as,for example, sodium, potassium, ammonium, calcium, or ferric hydroxides,and such organic bases as isopropylamine, trimethylamine,2-ethylamino-ethanol, histidine, procaine, and the like. Such salts areformed as acid addition salts with any free cationic groups andgenerally are formed with inorganic acids such as, for example,hydrochloric, sulfuric, or phosphoric acids, or organic acids such asacetic, citric, p-toluenesulfonic, methanesulfonic acid, oxalic,tartaric, mandelic, and the like. Salts of the disclosure may includeamine salts formed by the protonation of an amino group with inorganicacids such as hydrochloric acid, hydrobromic acid, hydroiodic acid,sulfuric acid, phosphoric acid, and the like. Salts of the disclosurealso include amine salts formed by the protonation of an amino groupwith suitable organic acids, such as p-toluenesulfonic acid, aceticacid, and the like. Additional excipients which are contemplated for usein the practice of the present disclosure are those available to thoseof ordinary skill in the art, for example, those found in the UnitedStates Pharmacopoeia Vol. XXII and National Formulary Vol. XVII, U.S.Pharmacopoeia Convention, Inc., Rockville, Md. (1989), the relevantcontents of which is incorporated herein by reference.

The “semisolid gel” according to the current disclosure is a semisolid.The semisolid formulation apparent viscosity may increase withconcentration.

As used herein, “sequential administration” includes that theadministration of two agents (e.g., compositions described herein)occurs separately on the same day or do not occur on a same day (e.g.,occurs on consecutive days).

The term, “serum” as used herein is the component that is neither ablood cell (serum does not contain white or red blood cells) nor aclotting factor. Serum is a protein-rich liquid that separates out whenblood coagulates; i.e., serum refers to components wherein fibrinogens,clotting factors, etc...are removed from the blood plasma. Serumincludes all proteins not used in blood clotting and all theelectrolytes, antibodies, antigens, hormones, and any exogenoussubstances (e.g., drugs and microorganisms). Blood is centrifuged toremove cellular components. Anti-coagulated blood yields plasmacontaining fibrinogen and clotting factors. Coagulated blood yieldsserum without fibrinogen, although some clotting factors remain. Inembodiments, “serum” is an engineered composition including componentsof natural human serum.

“Solution” according to the current disclosure is a clear, homogeneousliquid dosage form that contains one or more chemical substancesdissolved in a solvent or mixture of mutually miscible solvents. Asolution is a liquid preparation that contains one or more dissolvedchemical substances in a suitable solvent or mixture of mutuallymiscible solvents. Because molecules of a drug substance in solution areuniformly dispersed, the use of solutions as dosage forms generallyprovides assurance of uniform dosage upon administration and goodaccuracy when the solution is diluted or otherwise mixed.

The term “solvent,” as used herein, refers to a liquid solvent eitheraqueous or non-aqueous. The selection of the solvent depends notably onthe solubility of the composition on said solvent and on the mode ofadministration. Aqueous solvent may consist solely of water, or mayconsist of water plus one or more miscible solvents, and may containdissolved solutes such as sugars, buffers, salts or other excipients.The more commonly used non-aqueous solvents are the short-chain organicalcohols, such as, methanol, ethanol, propanol, short-chain ketones,such as acetone, and poly alcohols, such as glycerol. As used herein,serum may be used as a solvent for reconstitution of the freeze-driedcomposition described herein.

“Suspension” as used herein is a liquid dosage form that contains solidparticles dispersed in a liquid vehicle.

As used herein, the term “syndrome” may refer to a group of symptomsthat consistently occur together or a condition characterized by a setof associated symptoms. A syndrome (e.g., dry eye syndrome) may be a setof medical signs and symptoms that are correlated with each other andoften, are correlated with a specific disease. A disease on the otherhand, may be a health condition that has a clearly defined reason behindit. A syndrome (from the Greek word meaning ‘run together’) however, mayproduce a number of symptoms without an identifiable cause. They maysuggest the possibility of an underlying disease or even the chances ofdeveloping a disease.

“Tear breakup time” or “TBUT” or “tear film breakup time” or “TFBUT” asused herein may refer to a clinical test that measures the intervalbetween the individual’s last complete blink and the breakup of the tearfilm. The test may be used to assess for dry eye syndrome. To measureTBUT, fluorescein is instilled into the patient’s tear film and thepatient is asked not to blink while the tear film is observed under abroad beam of cobalt blue illumination. The TBUT is recorded as thenumber of seconds that elapse between the last blink and the appearanceof the first dry spot in the tear film.

The terms “treat,” “treating” or “treatment,” and other grammaticalequivalents as used herein, include alleviating, abating, ameliorating,or preventing a disease, condition or symptoms, preventing additionalsymptoms, ameliorating or preventing the underlying metabolic causes ofsymptoms, inhibiting the disease or condition, e.g., arresting thedevelopment of the disease or condition, relieving the disease orcondition, causing regression of the disease or condition, relieving acondition caused by the disease or condition, or stopping the symptomsof the disease or condition, and are intended to include prophylaxis.The terms further include achieving a therapeutic benefit and/or aprophylactic benefit. By therapeutic benefit is meant eradication oramelioration of the underlying disorder being treated. Also, atherapeutic benefit is achieved with the eradication or amelioration ofone or more of the physiological symptoms associated with the underlyingdisorder such that an improvement is observed in the patient,notwithstanding that the patient may still be afflicted with theunderlying disorder.

The term “umbilical cord blood” or “cord blood” as used herein generallyrefers to blood obtained from the placenta and the umbilical cord afterchild birth. This blood originates from the neonate and is no longerneeded by the newborn and is commonly discarded, for example, cord bloodmay refer to blood which is obtained from the umbilical cord or placentaof newborns. The use of cord or placental blood can be obtainedrelatively easily and without trauma to the donor. Cord blood ispreferably obtained by direct drainage from the umbilical vein of adiscarded placenta. For example, the umbilical cord blood may beobtained from mothers with vaginal or cesarean section delivery.

The term “umbilical cord blood serum” or “cord serum” generally refersto umbilical cord blood in which the cells have been removed so that thecord serum is substantially free of whole cells. Umbilical cord bloodserum contains a high concentration of tear components, growth factors,neurotrophic factors, vitamin A, fibronectin, prealbumin, and oil.Umbilical cord serum can provide basic nutrients for epithelial renewaland can facilitate the proliferation, migration, and differentiation ofthe ocular surface epithelium.

The term “vaginal ring” as used herein may refer to a thin and/orflexible ring that is inserted into the vagina. In some examples, theactive agent (e.g., reconstituted freeze-dried composition describedherein) may be controlled-release/sustain-release.

The term “vial” or “container”, as used herein, refers broadly to areservoir suitable for retaining the plasma or serum composition inlyophilized form. Similarly, it will retain the solvent forreconstitution. Examples of a vial that can be used in the presentdisclosure include syringes, ampoules, cartridges, or other suchreservoir suitable for delivery of the plasma or serum composition tothe patient. Alternatively, the vial retaining the plasma or serumcomposition the one retaining the solvent for reconstitution can bepresented as the 2 compartments of a dual-chamber system (syringe orcartridge for example). Vials suitable for packaging products forophthalmic administration are well known and recognized in the art.

As used herein, “viscosity” refers to a fluid’s resistance to flow.

The term “weight percent” or “% (w/w)” refers to a percentage of acomponent in a solution that is calculated on the basis of weight forthe component and the solvent. For example, a 1% (w/w) solution of acomponent would have 1 g of the component dissolved in a 100 g ofsolvent. The term “volume percent” or “% (v/v)” refers to a percentageof a component in a solution that is calculated on the basis of volumefor the component and the solvent. For example, a 1% (v/v) solution of acomponent would have 1 ml of the component dissolved in a 100 ml ofsolvent. The term “weight/volume percent” or “% (w/v)” refers to apercentage of a component in a solution that is calculated on the basisof weight for the component and on the basis of volume for the solvent.For example, a 1.0% (w/v) solution of a component would have 1 g of thecomponent dissolved in a 100 ml of solvent.

Composition

In one aspect, the present disclosure includes a freeze-driedcomposition including non-autologous plasma or serum and a polymer. Inembodiments, the present disclosure includes a freeze-dried compositionfor treating dry eye syndrome or for moisturizing and/or repairingnon-keratinized surfaces (e.g., dry mouth syndrome, vaginal dryness,diabetic ulcers and other chronic wounds), including non-autologousplasma or serum, and a polymer having a molecular weight of 100 to 1200kDA, or derivatives thereof.

In embodiments, the present disclosure includes a freeze-driedcomposition including non-autologous serum and a polymer for treatingdry eye syndrome or for moisturizing and/or repairing non-keratinizedsurfaces (e.g., dry mouth syndrome, vaginal dryness, diabetic ulcers andother chronic wounds), wherein the polymer has a molecular weight of 100to 1200 kDA, or derivatives thereof.

In embodiments, the serum or plasma is freeze-dried together with thepolymer to form a freeze-dried composition. In alternative embodiments,the serum or plasma can be freeze-dried separately from the polymer,which may or may not also be freeze-dried.

In embodiments, the disclosure provides a freeze-dried compositioncomprising non-autologous plasma or serum and a polysaccharide having amolecular weight of 100 to 1200 kDa, preferably from 100 to 400 kDa, ora polysaccharide having a molecular weight of 100, 250, or 400 kDa,wherein the polysaccharide is chitosan, or a hydrophobically modifiedchitosan. In embodiments, the composition preferably comprisesnon-autologous serum. In embodiments, the freeze-dried composition isbiodegradable and biocompatible. In embodiments, the freeze-driedcomposition is in the form of a cohesive solid material, such as a waferor sheet. In embodiments, the freeze-dried composition in the form of acohesive solid material is further combined with a sufficient amount ofan aqueous liquid (e.g., a reconstitution fluid). In embodiments, thereconstitution fluid is any solvent described herein. In exemplaryembodiments, water or a saline solution may be used as thereconstitution fluid, any additives may be included (e.g., glycine orascorbic acid), and in in additional embodiments, liquid serum may alsobe used as a reconstitution fluid to form a cohesive sponge-like solidmaterial. In embodiments where liquid serum or plasma is used, the serumor plasma may optionally be diluted with an acceptable carrier, such aswater or an aqueous buffer, e.g., saline. In another embodiment, thefreeze-dried composition in the form of a powder which can be formulatedinto a suitable dosage form for application to a non-keratinizedepithelial surface, such as the surface of the eye, mouth, or vagina, orthe surface of an external wound. Such powder compositions may beformulated, for example, as a liquid, gel, or ointment.

In one aspect, the present disclosure includes a freeze-driedcomposition for treating dry eye syndrome or for moisturizing and/orrepairing non-keratinized surfaces (e.g., dry mouth syndrome, vaginaldryness, diabetic ulcers and other chronic wounds), includingnon-autologous plasma or serum, and a polymer having a molecular weightof 100 to 1200 kDA and a formula including of Formula 1:[[R₁]m-L-[R₂]n]p, or derivatives thereof, where R₁ and R₂ are units ofthe polymer, m is an integer from 1-10,000, n is an integer from1-10,000, and p is an integer from 10-10,000, L is a bond. In someembodiments, “m” and “n” of Formula I is 1.

In embodiments, freeze-dried composition includes umbilical cord plasmaor serum. In embodiments, the polymer in the composition is a highmolecular weight (e.g. 100 to 1200 kDA) polymer such that thecomposition upon administration to the eye is retained on the ocularsurface due to the impermeability of the polymer through one or morelayers of the cornea, conjunctiva, and sclera. In embodiments, thepolymer included in the composition is chitosan. The polymer increasesthe ocular surface retention of the composition such that uponadministration to an eye of a subject having dry eye syndrome, thecomposition is retained on the ocular surface for about 4 hours to about24 hours.

In embodiments, the present disclosure includes a freeze-driedcomposition for treating dry eye syndrome or for moisturizing and/orrepairing non-keratinized surfaces (e.g., dry mouth syndrome, vaginaldryness, diabetic ulcers and other chronic wounds), includingnon-autologous plasma or serum, and chitosan or derivatives thereof. Inembodiments, the present disclosure includes a freeze-dried compositionfor treating dry eye syndrome or for moisturizing and/or repairingnon-keratinized surfaces (e.g., dry mouth syndrome, vaginal dryness,diabetic ulcers and other chronic wounds), including mammalian (e.g.,human) umbilical cord plasma and chitosan or derivatives thereof. Inembodiments, the present disclosure includes a freeze-dried compositionfor treating dry eye syndrome or for moisturizing and/or repairingnon-keratinized surfaces (e.g., dry mouth syndrome, vaginal dryness,diabetic ulcers and other chronic wounds), including mammalian (e.g.,human) umbilical cord serum and chitosan or derivatives thereof. In someembodiments, the non-autologous plasma or serum includes allogenic orhomologous plasma or serum, respectively.

Blood plasma or serum used in the composition of the present disclosuremay include plasma or serum isolated from blood of all species ofmammals including humans, sheep, goats, pigs, horses, dogs and cattle,primates, and rodents. In an embodiment of the present disclosure, bloodplasma or serum used in the composition can be purchased from a BloodBank. In some embodiments, the non-autologous plasma or serum of thepresent disclosure may be from umbilical cord plasma or serum.

In embodiments, the composition includes plasma or serum (e.g., humanumbilical cord plasma or serum) from about 0.001 percent by weight toabout 90 percent by weight of the composition, from about 0.001 percentby weight to about 1 percent by weight, from about 0.001 percent byweight to about 10 percent by weight, from about 0.001 percent by weightto about 20 percent by weight, from about 0.001 percent by weight toabout 30 percent by weight, from about 0.001 percent by weight to about40 percent by weight, from about 0.001 percent by weight to about 50percent by weight, from about 0.001 percent by weight to about 60percent by weight, from about 0.001 percent by weight to about 70percent by weight, from about 0.001 percent by weight to about 80percent by weight, from about 0.01 percent by weight to about 90 percentby weight, from about 0.01 percent by weight to about 1 percent byweight, from about 0.01 percent by weight to about 10 percent by weight,from about 0.01 percent by weight to about 20 percent by weight, fromabout 0.01 percent by weight to about 30 percent by weight, from about0.01 percent by weight to about 40 percent by weight, from about 0.01percent by weight to about 50 percent by weight, from about 0.01 percentby weight to about 60 percent by weight, from about 0.01 percent byweight to about 70 percent by weight, from about 0.01 percent by weightto about 80 percent by weight, from about 0.1 percent by weight to about90 percent by weight, from about 0.1 percent by weight to about 1percent by weight, from about 0.1 percent by weight to about 10 percentby weight, from about 0.1 percent by weight to about 20 percent byweight, from about 0.1 percent by weight to about 30 percent by weight,from about 0.1 percent by weight to about 40 percent by weight, fromabout 0.1 percent by weight to about 50 percent by weight, from about0.1 percent by weight to about 60 percent by weight, from about 0.1percent by weight to about 70 percent by weight, from about 0.1 percentby weight to about 80 percent by weight of the composition, and anyrange in between.

In embodiments, the freeze-dried composition contains from about 10-90wt% plasma or serum (e.g., human umbilical cord plasma or serum)components, or from 10-90 wt%, 20-90 wt%, 30-90 wt%, 40-90 wt%, 50-90wt%, 60-90 wt% plasma or serum components. In embodiments, thecomposition contains about 40 wt%, about 50 wt%, about 60 wt%, about 70wt%, or about 80 wt% plasma or serum components. In an embodiment, thefreeze-dried composition contains about 75 wt% plasma or serumcomponents. In the context of these embodiments, preferably theremainder of the weight percentage of the composition is made up of apolysaccharide, preferably chitosan or a hydrophobically modifiedchitosan. In certain embodiments, the composition may further containsmall amounts of additional additives or excipients, typically in arange of less 10 wt% collectively, preferably less than 5 wt% or lessthan 2 wt%, based on the total dry weight of the composition.

In embodiments, the composition of the present disclosure includes oneor more polymers. For example, the polymer included in the compositionincludes a low molecular weight polymer, a medium molecular weightpolymer, or a high molecular weight polymer. Low molecular weightpolymers may include for example, polymers of approximately 100 kDa orless (e.g., approximately 10 kDa to approximately 100 kDa). Mediumweight polymers may include, for example, polymers of approximately 250kDa or between 100 and 350 kDa, whereas high molecular weight polymersmay include polymers of about 400 kDa or from about 400 to 1200 kDa. Inembodiments, the composition of the present disclosure includesultra-high weight polymers, and may include polymers of greater than1000 kDa. In embodiments, the molecular weight of the polymer is in arange from about 100 kDa to about 1200 kDa, from about 100 kDa to about200 kDa, from about 100 kDa to about 300 kDa, from about 100 kDa toabout 400 kDa, from about 100 kDa to about 500 kDa, from about 100 kDato about 600 kDa, from about 100 kDa to about 700 kDa, from about 100kDa to about 800 kDa, from about 100 kDa to about 900 kDa, from about100 kDa to about 1000 kDa, from about 100 kDa to about 1100 kDa, fromabout 100 kDa to about 1200 kDa, and any weight there between.

In embodiments, the polymer is a chitosan and has a molecular weight offrom 100 to 400 kDa, or a molecular weight of 100, 250, or 400 kDa.

In embodiments, the polymer is a biopolymer, e.g., cellulose, starch,lignin, chitin, and various polysaccharides. In embodiments, the polymeris a polycationic polymer with positive charges at several sites, forexample, along the backbone of chitosan and its derivatives. Inembodiments, the polymer is chitosans, alginates, gelatins, or anycombination(s) thereof. In embodiments, the polymer is, for example, oneor more hydrophobically modified polysaccharides.

In embodiments, the polymer (e.g., chitosans, alginates, or gelatins) isadded to the composition in any suitable amounts. For example, in someembodiments, polymer is in a concentration of from about 0.025% to about2.0% by weight of the composition. The polymer can be present at about0.025, about 0.05, about 0.1, about 0.2, about 0.3, about 0.4, about0.5, about 1.0, and about 2.0 percent by weight or any amount in betweenthese amounts of the composition. In an embodiment, the polymer is addedat a concentration of about 0.05% (w/v) to about 1.0% (w/v) of thecomposition. In embodiments, the ratio of the non-autologous plasma orserum to the polymer in the composition is 1:1, 1.1:1, 1.2:1, 1:3:1,1:4:1, 1.5:1, 1.6:1, 1.7:1, 1.8:1, 1.9:1, 2.0:1, 2.1:1, 2.2:1, 2.3:1,2.4:1, 2.5:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, or any ratiointegers in between, and/or vice versa.

In embodiments, the polymer of the present composition may includechitosan or derivatives thereof. For example, the composition includessalts or derivatives of chitosan. Salts of chitosan include organic andinorganic acid salts, for example, glutamate, lactate, citrate,hydrochloride, succinate, maleate, ascorbate, propionate, formate,carbonate and acetate salts. Derivatives of chitosan includehydrophobically-modified chitosan, succinyl chitosan, carboxymethylchitosan, glycol chitosan, sulfonated chitosan, thiolated chitosan orchitosan modified with quaternary ammonium groups.

In embodiments, the polymer is a chitosan, and in certain embodiments,the chitosan is a hydrophobically-modified (hm) chitosan. A chitosan canbe hydrophobically-modified using standard techniques in the art. Incertain aspects, chitosans can be hydrophobically-modified by reactionof alkyl (or aryl) aldehydes with primary amine groups along thechitosan backbone (i.e., in a 50/50 (v/v)% of aqueous acetic acid andethanol). After reaction, the resulting Schiff bases, or imine groups,can be reduced to stable secondary amines by dropwise addition of areducing agent. In other aspects, the chitosan can behydrophobically-modified through the addition of palmitic anhydride(i.e., using 0.1 g dissolved into 20 mL of ethanol) to the chitosan(i.e., wherein the chitosan is dissolved in a 50/50 (v/v) mixture ofacetic acid).

In embodiments, the hydrophobic substituents of the hydrophobicallymodified chitosan are provided by palmitic anhydride. In embodiments,the disclosure provides a freeze-dried composition comprising plasma orserum and a polysaccharide selected from chitosan, a hydrophobicallymodified chitosan, sucrose, or trehalose, wherein the compositioncomprises from about 60-90 wt% of the plasma or serum components andfrom about 10-40 wt% of the polysaccharide.

In embodiments of the freeze-dried compositions described here in whichthe chitosan is a hydrophobically modified chitosan, the chitosan may behydrophobically-modified by reaction of alkyl (or aryl) aldehydes withprimary amine groups along the chitosan backbone in a 50/50 (v/v)% ofaqueous 0.2 M acetic acid and ethanol, as described in US 20160206777.As described in US 20160206777, after reaction, the resulting Schiffbases, or imine groups, are reduced to stable secondary amines bydropwise addition of the reducing agent sodium cyanoborohydride.Hydrophobically modified chitosans, and methods for making them, arealso described in U.S. Pat. Nos 9,616,088 and 8,932,560, the disclosuresof which are hereby incorporated by reference.

In embodiments of the freeze-dried compositions described here in whichthe chitosan is a hydrophobically modified chitosan, up to 10% of theavailable amines of the chitosan backbone are substituted with thehydrophobic substituent. In embodiments, between 1.5% and 4.5% ofavailable amines are substituted.

In embodiments, the hydrophobically-modified chitosan of the compositiondescribed here is selected from the group consisting of chitosanlactate, chitosan salicylate, chitosan pyrrolidone carboxylate, chitosanitaconate, chitosan niacinate, chitosan formate, chitosan acetate,chitosan gallate, chitosan glutamate, chitosan maleate, chitosanaspartate, and chitosan glycolate.

In embodiments, the polymer of the present composition is apolysaccharide selected from the group consisting of chitosan,cellulose, dextrin, pectin, alginic acid, agar, agarose, and carragenas,and derivatives thereof. In embodiments, the polysaccharide is chitosan,or a derivative thereof, preferably a hydrophobically modified chitosan.Chitosan is enzymatically degraded in vivo by several enzymes with themain enzyme being lysozyme which is a non-specific protease present inall mammalian tissues. Human lysozyme is part of the human body’sdefense against some bacteria. Chitosan is enzymatically degraded bylysozyme into non-toxic oligosaccharides that can be either excreted orincorporated into glycosoaminoglycans and glycoproteins. Lysozyme ispresent in almost all body and maternal fluids including blood serum,umbilical cord blood serum, amniotic fluid, saliva, gastric juice, tearsand mucous membranes. Activity is normally from 7 to 13 mg/L in serumand 120 times higher in tears. Gastric juice has about 8 times more thanin normal serum. Chitosan is both biodegradable and biocompatible.Chitosan is also chemically versatile and mimics the extracellularmatrices of tissues that are made up of various glycosaminoglycans. Thismakes chitosan and its derivatives especially advantageous for tissueengineering and drug delivery due to their ability to act as molecularscaffolds. Drugs within the chitosan matrix can be slowly released tothe diseased target as the chitosan is biodegraded by endogenouslysozyme and other enzymes.

Exemplary beneficial properties of chitosan and its derivatives includeanti-microbial properties, biodegradability, biocompatibility, andnontoxic properties. Furthermore, the mucoadhesive properties andviscosity of chitosan or derivatives thereof may readily be modified,for example by choice of a low, medium, or high molecular weightchitosan as described herein. Generally, as described in the examples,both mucoadhesiveness and viscosity of the polymer-containingcompositions of the disclosure increase with increasing molecular weightof the chitosan polymer. The molecular weight of chitosan can varybetween 10-100,000 kDa. Preferably, the compositions described herecontain chitosan, or a hydrophobically modified chitosan of a molecularweight ranging from 100 to 400 kDa, or 100, 250, or 400 kDa. Theanti-microbial properties of chitosan and its derivatives may be againstbacteria, yeast, fungi, and viruses. The anti-microbial effect isbelieved to be chitosan’s ability to breakdown the microorganism wall,thereby causing it to leak and eventually killing the microorganism.

As described above, the mucoadhesive properties and viscosity ofchitosan may be readily modified. Chitosan is a linear polysaccharidecomposed of linked glucosamine units. The ability to change themolecular structure of chitosan by varying degrees allows its propertiesto be modified. One property of chitosan is the immense structuralpossibilities for chemical and mechanical modifications to generatenovel properties, functions and applications. Even though molecularweight has a profound effect on the properties of chitosan,deacetylation, or the addition of different functional groups can alsochange its properties, e.g., depolymerized chitosans are morewater-soluble.

The chitosan has the following backbone chemical structure, where nrepresents any integer and represents the number of monomeric units inthe chitosan chain (i.e., the degree of polymerization).

In embodiments, the composition of the present disclosure includeschitosan derivatives, for example, but is not limited to, esters, ethersor other derivatives formed by bonding acyl and/or alkyl groups with thehydroxyl groups. Additional examples of chitosan derivatives includeO-alkyl ethers of chitosan and O-acyl esters of chitosan. Inembodiments, the composition of the present disclosure includes modifiedchitosans, such as those conjugated to polyethylene glycol may be usedin the present disclosure. Conjugates of chitosan and polyethyleneglycol are described in WO99/01498 and incorporated herein by reference.In embodiments, the composition of the present disclosure includeschitosan derivatives, for example, N-pathaloylation of chitosan,dendronized chitosan-sialic acid hybrids, methylthiocarbamoyl andphenylthiocarbamoyl chitosans, lactic/glycolic acid chitosan hydrogels,or nanocomposite from natural polysaccharide chitosan.

The chitosan, chitosan derivative, or salt in the composition of thepresent disclosure may have a molecular weight in the range a fewglucosamine units of to more than 200,000 Da. In embodiments, themolecular weight can range from 10,000 to 1,000,000 Da, in the range15,000 to 750,000 Da, or in the range 20,000 to 500,000 Da. Inembodiments, the chitosan may range in molecular weight from 300 to 1000kDa. In embodiments, the chitosan may range in molecular weight fromabout 300 to 400 kDa, or from about 300 to 500 kDa, or from about 300 to400 kDa, or from about 300 to 500 kDa, or from about 300 to 600 kDa, orfrom about 300 to 700 kDa, or from about 300 to 800 kDa, or from about300 to 900 kDa, or from about 300 to 1000 kDa.

In embodiments, chitosan is one of the polysaccharides that is modifiedwith various groups such as 5β-cholanic acid, linoleic acid, Monomethoxypoly (ethyleneglycol). After modification process, modified chitosan areused for preparation of the composition of the present disclosure.Non-limiting examples of modified chitosan and methods of preparing suchmodified chitosan are listed in Table 1.

TABLE 1 Functional molecules for modification of chitosan ChitosanGrafting Agent Glycol chitosan 5β-Cholanic acid Modification: Glycolchitosan is hydrophobically modified with cholanic acid inmethanol/water. To activate the carboxylic acid groups of cholanic acid,equal amounts of 1-ethyl-3-(3-dimethylaminopropyl) - carbodiimideydrochloride and N-hydroxysuccinimide are added. Chitosan of 100 meshLinoleic acid (LA) Modification: Chitosan is dissolved in aqueous aceticacid solution and diluted of methanol. LA is added to the chitosansolution glucosamine residue of chitosan followed by a dropwise additionof 15 mL of EDC methanol solution (0.07 g/L) while stirring. chitosanα-Cyclodextrin Modification: α-CD linked chitosan using2-O-formylmethyl-α-CD by reductive N-alkylation. Chitosan

Modification: The PCL-graft-chitosan copolymers are synthesized bycoupling the hydroxyl end-groups on preformed PCL chains and the aminogroups present on 6-Otriphenylmethyl chitosan and by removing theprotective 6-O-triphenylmethyl groups in acidic aqueous solutionBiomedical grade chitosan Monomethoxy poly(ethyleneglycol) Modification:Chitosan is completely dissolved in formic acid by stirring and asuitable amount of mPEG is added. After 15 min, enough formaldehydesolution is added to the above mixture and was stirred for 12 h.

In embodiments, the present disclosure includes a freeze-driedcomposition for treating dry eye syndrome or for moisturizing and/orrepairing non-keratinized surfaces (e.g., dry mouth syndrome, vaginaldryness, diabetic ulcers and other chronic wounds), includingnon-autologous plasma or serum, and a polymer having a molecular weightof 100 to 1200 kDA, or derivatives thereof, and one or more additives.The additives can include components that have been classified by theFDA as Ophthalmic Drug Products for Over-The-Counter Human Use [Title21, Volume 5]. In embodiments, these additives may be (a) astringents(b) buffering agents (c) demulcents (d) emollients (e) eyewash, eyelotion, irrigating solutions (f) hypertonic agents (g) isotonic agents,and/or (h) vasoconstrictors. In embodiments, these additives may includeZinc sulfate, 0.25 percent; cellulose derivatives:Carboxymethylcellulose sodium, 0.2 to 2.5 percent Hydroxyethylcellulose, 0.2 to 2.5 percent Hypromellose, 0.2 to 2.5 percentMethylcellulose, 0.2 to 2.5 percent, Dextran 70, 0.1 percent, Gelatin,0.01 percent; polyols liquid: Glycerin, 0.2 to 1 percent, Polyethyleneglycol 300, 0.2 to 1 percent, Polyethylene glycol 400, 0.2 to 1 percent,Polysorbate 80, 0.2 to 1 percent, Propylene glycol, 0.2 to 1 percent,Polyvinyl alcohol, 0.1 to 4 percent, Povidone, 0.1 to 2 percent; Lanolinpreparations: Anhydrous lanolin, 1 to 10 percent Lanolin, 1 to 10percent; Oleaginous ingredients: Light mineral oil, Mineral oil,Paraffin, Petrolatum, White ointment, White petrolatum, White wax,Yellow wax; and/or Ophthalmic vasoconstrictors: Ephedrine hydrochloride,0.123 percent, Naphazoline hydrochloride, 0.01 to 0.03 percent,Phenylephrine hydrochloride, 0.08 to 0.2 percent, Tetrahydrozolinehydrochloride, 0.01 to 0.05 percent.

In embodiments, the present disclosure includes a freeze-driedcomposition for treating dry eye syndrome or for moisturizing and/orrepairing non-keratinized surfaces (e.g., dry mouth syndrome, vaginaldryness, diabetic ulcers and other chronic wounds), includingnon-autologous plasma or serum, and a polymer having a molecular weightof 100 to 1200 kDA, or derivatives thereof, and trehalose,methylcellulose, ethylcellulose, sodium carboxymethylcellulose,hydroxypropylmethylcellulose, sodium hyaluronate, sodium alginate,chitosan, chitosan salts and/or derivatives, polyethylene glycol,glycerin, propylene glycol, Triacetin, N,N-Dimethylacetamide, poly(vinylpyrrolidone), pyrrolidone, dimethyl sulfoxide, ethanol,N-(-beta-Hydroxyethyl)-lactamide, 1-Methyl-2-pyrrolidinone,triglycerides, monothioglycerol, sorbitol, lecithin, methylparaben,propylparaben, or combinations thereof.

In embodiments, the present disclosure includes a freeze-driedcomposition for treating dry eye syndrome or for moisturizing and/orrepairing non-keratinized surfaces (e.g., dry mouth syndrome, vaginaldryness, diabetic ulcers and other chronic wounds), includingnon-autologous plasma or serum, and a polymer having a molecular weightof 100 to 1200 kDA, or derivatives thereof, and a co-solvent. Thesolubility of the components of the present compositions may be enhancedby a surfactant or other appropriate co-solvent in the composition. Suchco-solvents include, but are not limited to, polysorbate 20, polysorbate60, polysorbate 80, Pluronic F-68, Pluronic F-84 Pluronic P-103,cyclodextrin, and any other suitable agents, or a combination thereof.The co-solvents may be used in any suitable amounts. In embodiments,such co-solvents are used in a concentration of about 5% to about 50%(w/v) of the composition. In embodiments, a solvent that used in acomposition of the present disclosure is glycerol.

In embodiments, the present disclosure includes a freeze-driedcomposition for treating dry eye syndrome or for moisturizing and/orrepairing non-keratinized surfaces (e.g., dry mouth syndrome, vaginaldryness, diabetic ulcers and other chronic wounds), includingnon-autologous plasma or serum, and a polymer having a molecular weightof 100 to 1200 kDA, or derivatives thereof, and a preservative. Examplesof preservatives include an anti-microbial preservative, for example,benzalkonium chloride, thimerosal, chlorhexidine, chlorobutanol, methylparaben, propyl paraben, phenylethyl alcohol, edetate disodium sorbicacid, Onamer M Polyquat, cetyl bromide, cetyl pyridinium chloride,benzyl bromide, EDTA, phenylmercury nitrate, phenylmercury acetate,thimerosal, merthiolate, acetate and phenylmercury borate, polymyxin Bsulphate, methyl and propyl parabens, quaternary ammonium chloride,sodium benzoate, sodium proprionate, and sodium perborate, or anycombination(s) thereof.

In embodiments the preservatives may be used in any suitable amounts.For example, the antimicrobial and preservative may be used in an amountabout 0.001% by weight-1.0% by weight based on the total weight ofcomposition.

In embodiments, the present disclosure includes a freeze-driedcomposition for treating dry eye syndrome or for moisturizing and/orrepairing non-keratinized surfaces (e.g., dry mouth syndrome, vaginaldryness, diabetic ulcers and other chronic wounds), includingnon-autologous plasma or serum, and a polymer having a molecular weightof 100 to 1200 kDA, or derivatives thereof, and a viscosity agent. Anysuitable agent that can increase viscosity may be used. Viscosityincreased above that of simple aqueous solutions may be desirable toincrease ocular absorption of the active compound, to decreasevariability in dispensing the formulation, to decrease physicalseparation of components of a suspension or emulsion of the formulationand/or to otherwise improve the ophthalmic formulation. Such viscosityagents include, for example polyvinyl alcohol, polyvinyl pyrrolidone,methyl cellulose, hydroxy propyl methylcellulose, hydroxyethylcellulose, carboxymethyl cellulose, hydroxy propyl cellulose, otheragents known to those skilled in the art, or a combination thereof. Suchagents may be used in any suitable amounts.

The viscosity agents may be used in any suitable amounts. In one aspect,the viscosity agent may be employed at a level in a concentration offrom about 0.01% to about 3.0% by weight.

In embodiments, the present disclosure includes a freeze-driedcomposition for treating dry eye syndrome or for moisturizing and/orrepairing non-keratinized surfaces (e.g., dry mouth syndrome, vaginaldryness, diabetic ulcers and other chronic wounds), includingnon-autologous plasma or serum, and a polymer having a molecular weightof 100 to 1200 kDA, or derivatives thereof, and one or more pH adjustingagents. The pH adjustment agent may be, for example, sodium hydroxide,hydrochloric acid, citric acid, malic acid, tartaric acid, acetic acid,phosphoric acid, maleic acid, glycine, sodium lactate, lactic acid,sodium citrate, ascorbic acid, sodium acetate, acetic acid, sodiumbicarbonate, sodium carbonate, carbonic acid, sodium succinate, succinicacid, sodium benzoate, benzoic acid, sodium phosphates,tris(hydroxymethyl)aminomethane, histidine, histidine hydrochloride, orany combination(s) thereof.

Compounds useful as pH regulators include, but are not limited toinclude boric acid, sodium boric acid, sodium phosphate (including 1, 2and 3 basic phosphate, such as 1 basic sodium phosphate 1 hydrate, 2basic sodium phosphate 7 hydrate and mixtures thereof). Any other properbuffers can be used to stabilize the pH level of the ophthalmic liquidmedicine by conferring physiological pH approved for ophthalmic liquidmedicines. Since said buffers are just examples and these buffers arewell known in ophthalmologic field, a person skilled in the art canchoose proper buffers that can be used for the composition of thepresent disclosure.

In embodiments, the compositions of the present disclosure is formulatedas a solution, a suspension, a semi-solid gel, a gel, an emulsion,semi-liquid, an ointment, a cream, foam gel, or acontrolled-release/sustain-release vehicle. For example, the compositionmay be in the form of a contact lens solution, eyewash, eye drop, eyegel, eye ointment, and the like.

In embodiments, the freeze-dried composition is reconstituted with areconstitution fluid. The reconstitution fluid may refer to any solventdescribed herein. In embodiments, water or a saline solution may beused, any additives may be included into the reconstitution fluid, andartificial or non-autologous plasma or serum may also be used as areconstitution fluid. In embodiments, the solvents used for thereconstitution fluid is sterile.

In embodiments, the reconstitution fluid includes an additive (e.g.,ascorbic acid, glycine, or a combination thereof). In embodiments, anadditive included in the reconstitution fluid has been classified by theFDA as Ophthalmic Drug Products for Over-The-Counter Human Use [Title21, Volume 5]. In embodiments, these additives can be (a) astringents(b) buffering agents (c) demulcents (d) emollients (e) eyewash, eyelotion, irrigating solutions (f) hypertonic agents (g) isotonic agents,and/or (h) vasoconstrictors. In embodiments, these additives may includeZinc sulfate, 0.25 percent; cellulose derivatives:Carboxymethylcellulose sodium, 0.2 to 2.5 percent Hydroxyethylcellulose, 0.2 to 2.5 percent Hypromellose, 0.2 to 2.5 percentMethylcellulose, 0.2 to 2.5 percent, Dextran 70, 0.1 percent, Gelatin,0.01 percent; polyols liquid: Glycerin, 0.2 to 1 percent, Polyethyleneglycol 300, 0.2 to 1 percent, Polyethylene glycol 400, 0.2 to 1 percent,Polysorbate 80, 0.2 to 1 percent, Propylene glycol, 0.2 to 1 percent,Polyvinyl alcohol, 0.1 to 4 percent, Povidone, 0.1 to 2 percent; Lanolinpreparations: Anhydrous lanolin, 1 to 10 percent Lanolin, 1 to 10percent; Oleaginous ingredients: Light mineral oil, Mineral oil,Paraffin, Petrolatum, White ointment, White petrolatum, White wax,Yellow wax; and/or Ophthalmic vasoconstrictors: Ephedrine hydrochloride,0.123 percent, Naphazoline hydrochloride, 0.01 to 0.03 percent,Phenylephrine hydrochloride, 0.08 to 0.2 percent, Tetrahydrozolinehydrochloride, 0.01 to 0.05 percent.

The freeze-dried composition of the disclosure can be reconstituted, forexample, under sterile condition, with a solvent, such as water or asaline solution (e.g. 0.9% w/v sodium chloride for injection) prior touse, thereby generating a reconstituted liquid or semi-liquidcomposition. In embodiments, freeze-dried serum and plasma arereconstituted in sterile water or other comparable solvents and approvedadditives to allow the ability to titrate pH, osmolarity, surfacetension, and viscosity. Because CO₂ is removed during the freeze driedprocess which results in an alkaline pH (8.2), the sterile watercontains additives including glycine (pH 2.4) and ascorbic acid toobtain a final pH of approximately 7.4. In some embodiments, plasma orserum is used as a reconstitution fluid for treatment of dry eyesyndrome, or other ophthalmic diseases because serum contains many keycomponents as does plasma, which include growth factors, vitamins,minerals and natural lubricants such as lipids. In embodiments, thereconstituted composition is tested in an animal model. For example, thecomposition including non-autologous plasma or serum and a polymer asdescribed herein can be used in an animal model to determine theefficacy, toxicity, or side effects of treatment with said composition.

Pharmaceutical Compositions and Formulations

The disclosure also provides pharmaceutical compositions includingnon-autologous serum and a polymer. In some embodiments, thepharmaceutical composition comprises non-autologous serum and a polymer,and at least one pharmaceutically acceptable excipient or carrier.Preferably, the amount is a therapeutically effective amount.

In embodiments the pharmaceutical composition may include apharmaceutically acceptable carrier. The type of carrier can be selectedbased upon the intended route of administration. Pharmaceuticallyacceptable carriers include sterile aqueous solutions (e.g., sterilewater) or dispersions and sterile powders for the extemporaneouspreparation of sterile topical solutions or dispersions. The use of suchmedia and agents for pharmaceutically active substances is well known inthe art. Except insofar as any conventional media or agent isincompatible with the composition (e.g., freeze-dried plasma or serum),use thereof in the freeze-dried compositions for the disclosure iscontemplated.

The term “pharmaceutically acceptable” refers to those compounds,materials, compositions, carriers, and/or dosage forms which are, withinthe scope of sound medical judgment, suitable for use in contact withthe tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

In embodiments, the pharmaceutical composition may include apharmaceutically acceptable excipient, which may refer to an excipientthat is useful in preparing a pharmaceutical composition that isgenerally safe, non-toxic and neither biologically nor otherwiseundesirable, and includes excipient that is acceptable for veterinaryuse as well as human pharmaceutical use. Examples of pharmaceuticallyacceptable excipients include, without limitation, polyols (for example,glycerol, propylene glycol, liquid polyethylene glycol and the like),oils, detergents, suspending agents, carbohydrates (e.g., glucose,lactose, sucrose or dextran), antioxidants (e.g., ascorbic acid orglutathione), chelating agents, low molecular weight proteins, orsuitable mixtures thereof. In embodiments, the excipients may beselected from sucrose and trehalose.

In embodiments, the pharmaceutical composition is in the form of aliquid or solution formulation suitable for topical administration as adrop, particularly as an ophthalmic drop. In embodiments, thepharmaceutical composition is in the form of a solid composition such asa powder suitable for reconstitution into a liquid form. In embodiments,the pharmaceutical composition is in the form of a solid or semi-solidcomposition such as an ointment, paste, cream, lotion, or gel. Inembodiments, the pharmaceutical composition is in the form of a cohesivesolid material in a defined shape, such as a wafer or disc(alternatively, a lens, pessary, or denture) which may be used as, or incombination with a dressing, e.g., a wound dressing. In embodiments, thepharmaceutical composition is formulated for topical administration byapplication to a non-keratinized surface such as an ophthalmic, oral,vaginal, buccal, sublingual, or rectal surface. For example, apharmaceutical composition of the invention may be in the form of anaqueous solution in the form of eye drops, or a solid in the form of adisc, wafer, or lens, e.g., a contact lens, a pessary, a wound dressing,or denture. In embodiments, the pharmaceutical composition is sterile.

Methods of Treatment or Use

In one aspect, the present disclosure provides a method of treating anophthalmic disease of the anterior segment (e.g., dry eye syndrome) in asubject in need thereof, the method including administering to an eye ofthe subject a reconstituted freeze-dried composition includingnon-autologous plasma or serum and a polymer, in an amount to treat dryeye syndrome in the subject. In embodiments, the polymer or derivativesthereof has a molecular weight of 100 to 1200 kDA.

In one aspect, the present disclosure provides a method of moisturizingand/or repairing non-keratinized surfaces (e.g., dry mouth syndrome,vaginal dryness, diabetic ulcers and other chronic wounds) in or on thebody of a subject. The method of moisturizing and/or repairingnon-keratinized surfaces (e.g., dry mouth syndrome, vaginal dryness,diabetic ulcers and other chronic wounds) includes, e.g., delivering acomposition of the present disclosure with the therapeutic agentdelivery device (100) of the present disclosure to a non-keratinizedsurface of a subject in need thereof.

In embodiments, the moisturizing and/or repairing non-keratinizedsurfaces includes treatment of dry mouth syndrome, vaginal dryness,diabetic ulcers, and/or chronic wounds.

In one aspect, the present disclosure provides a method of treating anophthalmic disease of the anterior segment (e.g., dry eye syndrome) in asubject in need thereof, the method including administering to an eye ofthe subject a reconstituted freeze-dried composition includingnon-autologous plasma or serum and a polymer having a molecular weightof 100 to 1200 kDA and a formula including of Formula 1:[[R₁]m-L-[R₂]n]p, or derivatives thereof, where R₁ and R₂ are units ofthe polymer, m is an integer from 1-10,000, n is an integer from1-10,000, and p is an integer from 10-10,000, L is a bond. In someembodiments, “m” and “n” of Formula I is 1.

In one aspect, the present disclosure provides a method of increasingocular surface retention of an ophthalmic composition, the methodincluding administering to an eye of the subject a composition includingnon-autologous plasma or serum and a polymer having a molecular weightof 100 to 1200 kDA and/or a formula including of Formula 1:[[R₁]m-L-[R₂]n]p, or derivatives thereof, where R₁ and R₂ are units ofthe polymer, m is an integer from 1-10,000, n is an integer from1-10,000, and p is an integer from 10-10,000, L is a bond. In someembodiments, “m” and “n” of Formula I is 1.

In one aspect, the present disclosure provides a method of moisturizingand/or repairing non-keratinized surfaces (e.g., dry mouth syndrome,vaginal dryness, diabetic ulcers and other chronic wounds) in a subjectin need thereof, the method including administering to thenon-keratinized surface a composition including reconstitutedfreeze-dried composition including non-autologous plasma or serum and apolymer having a molecular weight of 100 to 1200 kDA and/or a formulaincluding of Formula 1: [[R₁]m-L-[R₂]n]p, or derivatives thereof, whereR₁ and R₂ are units of the polymer, m is an integer from 1-10,000, n isan integer from 1-10,000, and p is an integer from 10-10,000, L is abond. In some embodiments, “m” and “n” of Formula I is 1.

The freeze-dried composition of the present disclosure is reconstitutedwith a reconstitution fluid. The reconstitution fluid can be any aqueoussolvent. For example, distilled or sterile water or a saline solution(e.g. 0.9% w/v sodium chloride for injection). In embodiments, thereconstitution fluid includes additives (i.e., glycine or ascorbic acid,or chitosan, alginate, and gelatin). In some embodiments, thereconstitution fluid includes plasma or serum, or components thereof. Inembodiments, the solvents used for the reconstitution fluid may besterile.

In embodiments, the present disclosure includes a method of treating dryeye syndrome or a method of moisturizing and/or repairingnon-keratinized surfaces (e.g., dry mouth syndrome, vaginal dryness,diabetic ulcers and other chronic wounds) in a subject in need thereof,the method including administering to an eye or a non-keratinizedsurface of the subject, respectively, a reconstituted freeze-driedcomposition including non-autologous plasma or serum, and chitosan orderivatives thereof. In embodiments, the present disclosure includes amethod of treating dry eye syndrome or a method of moisturizing and/orrepairing non-keratinized surfaces (e.g., dry mouth syndrome, vaginaldryness, diabetic ulcers and other chronic wounds) in a subject in needthereof, the method including administering to an eye or anon-keratinized surface of the subject, respectively, a reconstitutedfreeze-dried composition including mammalian (e.g., human) umbilicalcord plasma and chitosan or derivatives thereof. In embodiments, thepresent disclosure includes a method of treating dry eye syndrome or amethod of moisturizing and/or repairing non-keratinized surfaces (e.g.,dry mouth syndrome, vaginal dryness, diabetic ulcers and other chronicwounds) in a subject in need thereof, the method including administeringto an eye or a non-keratinized surface of the subject, respectively, areconstituted freeze-dried composition including mammalian (e.g., human)umbilical cord serum and chitosan or derivatives thereof.

Treatment of Types of Dry Eye Syndrome

In embodiments, the method of treating dry eye syndrome includestreating aqueous tear-deficient dry eye, which is a disorder in whichthe lacrimal glands fail to produce enough of the watery component oftears to maintain a healthy eye surface, with a composition of thepresent disclosure. In embodiments, the method of treating dry eyesyndrome includes treating evaporative dry eye, which may result frominflammation of the meibomian glands (these glands make the lipid oroily part of tears that slows evaporation and keeps the tears stable),also located in the eyelids, with a composition of the presentdisclosure.

In embodiments, the method of treating dry eye syndrome includestreating dry eye associated with or resulting from treating inflammationof the surface of the eye, the lacrimal gland, or the conjunctiva; dryeye associated with any disease process that alters the components ofthe tears; dry eye associated with an increase in the surface of theeye, as in thyroid disease when the eye protrudes forward; and/or dryeye associated with a cosmetic surgery, for example, if the eyelids areopened too widely during surgery.

In embodiments, the method of treating dry eye syndrome includesameliorating a symptom of dry eye syndrome, including: stinging orburning of the eye; a sandy or gritty feeling as if something is in theeye; episodes of excess tears following very dry eye periods; a stringydischarge from the eye; pain and redness of the eye; episodes of blurredvision; heavy eyelids; inability to cry when emotionally stressed;uncomfortable contact lenses; decreased tolerance of reading, working onthe computer, or any activity that requires sustained visual attention;and/or eye fatigue.

In embodiments, the method of treating dry eye syndrome includestreating or ameliorating a symptom associated of dry eye syndrome in asubject exposed to a risk factor of temporary or chronic dry eye suchas: side effect of some medications, including antihistamines, nasaldecongestants, tranquilizers, certain blood pressure medicines,Parkinson’s medications, birth control pills and anti-depressants; skindisease on or around the eyelids can result in dry eye; diseases of theglands in the eyelids, such as meibomian gland dysfunction; pregnancy;hormone replacement therapy in women (women taking only estrogen are 70percent more likely to experience dry eye, whereas those taking estrogenand progesterone have a 30 percent increased risk of developing dryeye); refractive surgery known as LASIK; chemical and thermal burns thatscar the membrane lining the eyelids and covering the eye; allergies;infrequent blinking, associated with staring at computer or videoscreens; both excessive and insufficient dosages of vitamins that cancontribute to dry eye; homeopathic remedies that may have an adverseimpact on a dry eye condition; loss of sensation in the cornea fromlong-term contact lens wear can lead to dry eye; dry eye can beassociated with immune system disorders such as Sjögren’s syndrome,lupus, and rheumatoid arthritis (Sjögren’s leads to inflammation anddryness of the mouth, eyes, and other mucous membranes. It can alsoaffect other organs, including the kidneys, lungs and blood vessels);dry eye can be a symptom of chronic inflammation of the conjunctiva, themembrane lining the eyelid and covering the front part of the eye, orthe lacrimal gland (chronic conjunctivitis can be caused by certain eyediseases, infection, exposure to irritants such as chemical fumes andtobacco smoke, or drafts from air conditioning or heating); if thesurface area of the eye is increased, as in thyroid disease when the eyeprotrudes forward or after cosmetic surgery if the eyelids are openedtoo widely, dry eye can result; dry eye may occur from exposurekeratitis, in which the eyelids do not close completely during sleep.

In embodiments, the disclosure provides a method of treating dry eyesyndrome in a subject in need of such treatment by increasing the tearfilm breakup time in the eyes of the subject. Break-up of the tear filmis initiated at points where the surface of the corneal epithelium isirregular and where epithelial cells have been damaged or have losttheir wettability. Active components in the freeze-dried plasma or serumcompositions described here promote the health of the cornealepithelium, especially with improved stratification and differentiationof corneal epithelial cells that produce a surface glycocalyx. Inembodiments, the disclosure also provides methods of reducing,ameliorating, or preventing damage to the corneal epithelium, forexample as indicated by the uptake of Rose Bengal dye.

Co-Administration - With Other Drugs/Treatment Methods

In embodiments, the method of treating dry eye syndrome can be managedas an ongoing condition. In embodiments, whether there is a disease thatis the underlying cause of the dry eye (such as Sjögren’s syndrome orlacrimal and meibomian gland dysfunction) is determined. In embodiments,if there is an underlying disease, that disease is concurrently treated.

In embodiments, the method includes treating dry eye with a compositionof the present disclosure in combination with cyclosporine (cyclosporineis an antiinflammatory medication available to treat dry eye). Inembodiments, the method of treating dry eye syndrome includesadministering a composition including non-autologous plasma or serum anda polymer having molecular weight of 100 to 1200 kDA, or derivativesthereof, in combination with cyclosporine. In embodiments, the methodincludes administering a composition of the present disclosureconcurrently, sequentially, simultaneously with cyclosporine. Inembodiments, the method of treating dry eye syndrome includesadministering a composition including non-autologous plasma or serum anda polymer having molecular weight of 100 to 1200 kDA, or derivativesthereof, in combination with a corticosteroid eye drop for decreasinginflammation. In embodiments, the method includes administering acomposition of the present disclosure concurrently, sequentially,simultaneously with a corticosteroid. In embodiments, the methodincludes administering a composition of the present disclosureconcurrently, sequentially, simultaneously with cyclosporine and acorticosteroid.

In embodiments, the method of treating dry eye syndrome includesadministering a composition of the present disclosure to a subjectwearing contact lens.

In embodiments, the method of treating dry eye syndrome includesadministering a composition of the present disclosure to a subject inwhom another method has been performed to treat dry eye. For example, inone such method a physician has plugged the drainage holes or punctum,which are small circular openings at the inner corners of the eyelidswhere tears drain from the eye into the nose, before administering acomposition of the present disclosure. In embodiments, tear duct plugs,also called punctal plugs, has been inserted by an eye care professional(Punctal plugs are made of silicone or collagen, are reversible, and area temporary measure). In severe cases, permanent plugs have beeninserted before administering a composition of the present disclosure.

In additional embodiments, a simple surgery, called punctal cautery, hasbeen performed permanently close the drainage holes, beforeadministering a composition of the present disclosure. The procedurehelps keep the limited volume of tears on the eye for a longer period oftime.

In embodiments, the method of treating dry eye syndrome or the method ofmoisturizing and/or repairing non-keratinized surfaces (e.g., dry mouthsyndrome, vaginal dryness, diabetic ulcers and other chronic wounds)includes administering a composition of the present disclosure to asubject with dry eye or a non-keratinized surface, and the subject is onsupplements or dietary sources (such as salmon) of omega-3 fatty acids(especially DHA and EPA) for decreasing symptoms of irritationassociated with dry eye or a dry non-keratinized surface.

In embodiments, the method of moisturizing and/or repairingnon-keratinized surfaces (e.g., dry mouth syndrome, vaginal dryness,diabetic ulcers and other chronic wounds) can be managed as an ongoingcondition. In embodiments, the disease that is the underlying cause isdetermined. In embodiments, if there is an underlying disease, thatdisease is concurrently treated.

In embodiments, the method of moisturizing and/or repairingnon-keratinized surfaces (e.g., dry mouth syndrome, vaginal dryness,diabetic ulcers and other chronic wounds) includes administering acomposition including non-autologous plasma or serum and a polymerhaving molecular weight of 100 to 1200 kDA, or derivatives thereof, incombination with any suitable lubricating and/or moisturizing agents.Exemplary vaginal moisturizers include, but are not limited to,Replens®, K-Y Liquibeads®, Lubrin®, Astroglide Silken Secret® andVitamin E gel. Examples of oral moisturizes (i.e. for dry mouth) mayinclude, but are not limited to, artificial saliva products, salivastimulants, Salese Soothing®, Orajel®, and Eucerin® cream. Exemplarymoisturizers for diabetic ulcers and/or chronic wounds may include anysaline or similar dressings that provide a moist environment, TriDerma®MD Ulcer Healing Cream, or Neoteric Diabetic Healing Cream®.

In embodiments, the method of moisturizing and/or repairingnon-keratinized surfaces (e.g., dry mouth syndrome, vaginal dryness,diabetic ulcers and other chronic wounds) includes administering acomposition including non-autologous plasma or serum and a polymerhaving molecular weight of 100 to 1200 kDA, or derivatives thereof, incombination with any suitable lubricating and/or moisturizing agents. Inembodiments, the method includes administering a composition of thepresent disclosure concurrently, sequentially, and simultaneously withany suitable lubricating and/or moisturizing agents. In embodiments, themethod includes administering a composition of the present disclosureconcurrently, sequentially, and simultaneously with cyclosporine and anysuitable lubricating and/or moisturizing agents.

In embodiments, the method of moisturizing and/or repairingnon-keratinized surfaces (e.g., dry mouth syndrome, vaginal dryness,diabetic ulcers and other chronic wounds) includes administering acomposition of the present disclosure to a subject in whom anothermethod has been performed to moisturize and/or repair non-keratinizedsurfaces (e.g., dry mouth syndrome, vaginal dryness, diabetic ulcers andother chronic wounds).

In embodiments, the method of moisturizing and/or repairingnon-keratinized surfaces (e.g., dry mouth syndrome, vaginal dryness,diabetic ulcers and other chronic wounds) includes administering acomposition of the present disclosure to a subject with dry eye or anon-keratinized surface, and the subject is on supplements or dietarysources (such as salmon) of omega-3 fatty acids (especially DHA and EPA)for decreasing symptoms of irritation associated with a drynon-keratinized surface.

Dosage Regimen

In embodiments, the method of treating dry eye syndrome or moisturizingand/or repairing non-keratinized surfaces (e.g., dry mouth syndrome,vaginal dryness, diabetic ulcers and other chronic wounds) includesadministering a composition of non-autologous plasma or serum to asubject in any suitable or therapeutically effective amount, e.g., fromabout 0.001 percent by weight to about 90 percent by weight of thecomposition, from about 0.001 percent by weight to about 1 percent byweight, from about 0.001 percent by weight to about 10 percent byweight, from about 0.001 percent by weight to about 20 percent byweight, from about 0.001 percent by weight to about 30 percent byweight, from about 0.001 percent by weight to about 40 percent byweight, from about 0.001 percent by weight to about 50 percent byweight, from about 0.001 percent by weight to about 60 percent byweight, from about 0.001 percent by weight to about 70 percent byweight, from about 0.001 percent by weight to about 80 percent byweight, from about 0.01 percent by weight to about 90 percent by weight,from about 0.01 percent by weight to about 1 percent by weight, fromabout 0.01 percent by weight to about 10 percent by weight, from about0.01 percent by weight to about 20 percent by weight, from about 0.01percent by weight to about 30 percent by weight, from about 0.01 percentby weight to about 40 percent by weight, from about 0.01 percent byweight to about 50 percent by weight, from about 0.01 percent by weightto about 60 percent by weight, from about 0.01 percent by weight toabout 70 percent by weight, from about 0.01 percent by weight to about80 percent by weight, from about 0.1 percent by weight to about 90percent by weight, from about 0.1 percent by weight to about 1 percentby weight, from about 0.1 percent by weight to about 10 percent byweight, from about 0.1 percent by weight to about 20 percent by weight,from about 0.1 percent by weight to about 30 percent by weight, fromabout 0.1 percent by weight to about 40 percent by weight, from about0.1 percent by weight to about 50 percent by weight, from about 0.1percent by weight to about 60 percent by weight, from about 0.1 percentby weight to about 70 percent by weight, from about 0.1 percent byweight to about 80 percent by weight, or any range in between, of thecomposition.

In embodiments, a method of treating an ophthalmic disease of theanterior segment (e.g., dry eye syndrome) or a method of moisturizingand/or repairing non-keratinized surfaces (e.g., dry mouth syndrome,vaginal dryness, diabetic ulcers and other chronic wounds) in a subjectin need thereof, includes administering to an eye or a non-keratinizedsurface of the subject, respectively, a reconstituted freeze-driedcomposition including non-autologous plasma or serum and a polymer(e.g., chitosans, alginates, or gelatins). In embodiments, the amount ofpolymer in the composition is at a concentration of from about 0.025% toabout 2.0% by weight of the composition. The polymer can be present atabout 0.025, about 0.05, about 0.1, about 0.2, about 0.3, about 0.4,about 0.5, about 1.0, and about 2.0 percent by weight or any amount inbetween these amounts of the composition. In an embodiment, the polymermay be in a concentration of about 0.05% (w/v) to about 1.0% (w/v) ofthe composition.

In embodiments, a method of treating an ophthalmic disease of theanterior segment (e.g., dry eye syndrome) or a method of moisturizingand/or repairing non-keratinized surfaces (e.g., dry mouth syndrome,vaginal dryness, diabetic ulcers and other chronic wounds) in a subjectin need thereof, includes administering to an eye or a non-keratinizedsurface of the subject, respectively, a reconstituted freeze-driedcomposition including non-autologous plasma or serum and a polymer,formulated in the form of a solution, a suspension, a semi-solid gel, agel, an emulsion, semi-liquid, an ointment, a cream, foam gel, or acontrolled-release/sustain-release vehicle. For example, the compositionmay be in the form of a contact lens solution, eyewash, eye drop, eyegel, eye ointment, and the like.

In embodiments, a method of treating an ophthalmic disease of theanterior segment (e.g., dry eye syndrome) or a method of moisturizingand/or repairing non-keratinized surfaces (e.g., dry mouth syndrome,vaginal dryness, diabetic ulcers and other chronic wounds) in a subjectin need thereof includes topically administering to an eye or anon-keratinized surface of the subject, respectively, a reconstitutedfreeze-dried composition including non-autologous plasma or serum and apolymer. In embodiments, the reconstituted composition (e.g., ophthalmiccomposition) is administered in the form of eyedrops. In embodiments thereconstituted composition (e.g., ophthalmic composition) may beformulated as a solution, suspension, semi-liquid, semi-solid gel, gel,emulsion, ointment, or cream.

In embodiments, the reconstituted composition (e.g., ophthalmiccomposition) may be administered topically to an eye or anon-keratinized surface in a dose range from about 0.001 mg to about 100mg mg per eye or desired non-keratinized surface area, respectively. Insome embodiments, the dosage for one eye or a desired area of anon-keratinized surface may be about one drop of reconstitutedcomposition which may correspond to about 50 µL to about 150 µL ofreconstituted composition.

In embodiments, the reconstituted composition may be administeredtopically to an eye or a non-keratinized surface by placing one to twodrops or more in each eye or a desired non-keratinized surface area,respectively, 1 to 24 times daily. For example, the composition may beapplied, 1, 2, 3, 4, 8, 12, 18 or 24 times a day, or more. In anembodiment, the composition may be applied by placing one or two dropsin each eye or a desired area of a non-keratinized surface,respectively, once or twice daily.

In embodiments, the moisturizing of a mucosal surface increasing themoisture content of the surface by about 5% to about 100% (compared tophysiologically normal non-keratinized surface). In embodiments,repairing of a mucosal surface increasing the moisture content of thesurface by about 5% to about 100% (compared to physiologically normalnon-keratinized surface) and healing a wound (e.g., abrasions, cuts,ulcers, tissue damages).

Delivery Device

In one aspect, the current disclosure provides a therapeutic agentdelivery device (100) which incorporates the above compositionalelements (e.g., the freeze-dried non-autologous plasma or serum andpolymer, and a reconstitution fluid) in one or morecompartments/chambers (101 and/or 102). The delivery device (100)includes two-chambers (Chambers A (101) and B (102)). Chamber A (101) isconfigured to contain freeze-dried, sterile, freeze-dried plasma orserum. Chamber B (102) is configured to contain aqueous reconstitutionfluid including pH adjustment elements (e.g. ascorbic acid) and/or otheradditives as protectants (e.g. chitosan, alginate, gelatin). Anon-permeable membrane (103) between Chamber A (101) and Chamber B (102)is configured to separate chamber A (101) and chamber B (102).

The non-permeable membrane (103) can be broken on-demand by a user ofthe delivery device (100), in order to mix the lyophilized (orfreeze-dried) plasma or serum and the reconstitution fluid. Themechanism of membrane disruption can occur by several means: one exampleincludes a digital mechanism, by which the user presses a button on thedelivery device (100), and an electromechanical signal causes themembrane to become permeable; in another example, the delivery device(100) can be agitated or twisted by the user in order to disrupt themembrane and allow mixing of components in Chamber A (101) and Chamber B(102).

At the distal end of the delivery device (100) is a dropper (106), whichis configured to dispense the resulting fluid upon mixing of chamber A(101) and chamber B (102); the dropping of droplets may occur by theuser squeezing the body of the delivery device (100). This deliverydevice (100) can be stored at room temperature as the freeze-dried serumor plasma does not degrade at the speed it would in a wet state.

The therapeutic agent delivery device (100) is covered or surrounded bya chamber casing (104). The ophthalmic therapeutic agent delivery device(100) includes a dropper casing (105).

The therapeutic agent delivery device (100) has longitudinal dimensionof the device is about 5 cm - 10 cm. The ophthalmic therapeutic agentdelivery device (100) of claim 31, wherein the horizontal dimension ofthe device is about 1 cm - 4 cm.

In embodiments, the therapeutic agent delivery device (100) of thepresent disclosure is used for delivering a composition. For example,the therapeutic agent delivery device (100) includes a freeze-driedcomposition in chamber A (101) and upon reconstitution with the fluid inchamber B (102) is delivered to the eye of a subject through the dropper(106) for treating dry eye syndrome.

In embodiments, the therapeutic agent delivery device (100) of thepresent disclosure includes a freeze-dried composition in the firstchamber (101) and a reconstitution fluid in the second chamber (102),and upon reconstitution, the reconstituted composition is delivered to asubject.

In embodiments, the therapeutic agent delivery device (100) of thepresent disclosure includes the reconstituted composition which is fortreating dry eye syndrome, or moisturizing and/or repairing anon-keratinized surface.

EXAMPLES Example 1: Serum and Plasma Collection

Voluntary pedigree donors are kept in either single donor lots or as ABOtype specific minipool (< 11 donors) lots as a precaution againstpotential infectious agents including prion risk. Both serum and plasmaare from whole blood collections. Human umbilical cord serum and plasmaare from voluntary pedigree mothers during vaginal or cesarean sectiondelivery.

Blood is collected at FDA licensed, US collection centers, and totalquality system and good manufacturing practice (GMP) facilities areused. To ensure safety, the product undergoes two mandatory phases,incorporating safety measures inherent in FDA licensed plasma.

Example 2: Serum and Plasma Pathogen Reduction Phase 1 PathogenReduction: Pedigree Donors and Quarantine

Voluntary pedigree donors undergo physical examination and screening todetect the presence of infection in accordance with FDA guidelines.Exemplary infections tested for include, for example, Trypanosoma cruzi,Hepatitis B and C virus, Human Immunodeficiency Virus Types 1 and 2(HIV-1 and HIV-2), Treponema pallidum (TPHA), Toxoplasmosis (TOXO),Cytomegalovirus (CMV), Human T-Lymphotropic virus (HTLV-I/II) and WestNile virus (WNV). Multiplex nucleic acid testing is performed, on forexample, HIV, Hepatitis B Virus (HBV), and Hepatitis C Virus-nucleicacid testing (HCV-NAT). A hemovigilance program is utilized. Donor Serumand plasma is quarantined frozen before undergoing processing untilrepeat donor examinations and testing again at 4 months ensures noinfections are detected.

Phase 2 Pathogen Reduction

Serum and plasma undergoes pathogen reduction after completion of phase1, using the Intercept Blood System. The system uses Amotosalen HCI, asynthetic psoralen, or other similar method. Typically a photoactivesolvent/detergent is mixed into the serum or plasma and exposed toultraviolet light. With Amotosalen HCI, photoactivation occurs when themixture is illuminated with UVA treatment at 3 J/cm². Once treated, theserum and plasma are stored and frozen at minus 70° C. for futurefreezing drying.

Phase 3: Pathogen Reduction (Optional)

Gamma irradiation between 10 and 50 kGy is performed to the freeze-driedserum and plasma at 4° C. Ascorbate is added as a protectant for plasmaproteins against gamma irradiation and for pH balance.

Example 3: Freeze-Drying of Serum and Plasma

Samples are thawed and incubated at 23° C. for 1 hour, and centrifugedat 3,000 g for 10 minutes with retention of the resulting supernatant.300 mL aliquots of the serum/plasma supernatant for lyophilization areplaced in a sterile 1 L cylindrical Pyrex (10 cm diameter, 25 cm height)vessel, and frozen in an even layer on the bottom and side of thecontainer by slow rotation in isopropanol dry-ice slurry.

The resulting frozen serum/plasma is then freeze dried for 48 hours at8° C. in a Virus Unitop 600 SL lyophilizer or other comparable unit.

Additives in measured amounts are used for pH balance and as aprotectant for serum/plasma proteins against gamma irradiation. Thefreeze-dried serum/plasma can be stored at or below room temperature(25° C.). Temperatures of 50° C. or greater for storage because of riskof degradation of plasma/serum proteins.

Example 4: Reconstitution of Freeze-Dried Plasma/Serum

Freeze-dried serum and plasma are reconstituted in sterile water (i.e.in 300 mL aliquots), and optionally along with other comparable solventsand approved additives to allow the ability to titrate pH, osmolarity,surface tension, and viscosity. The sterile water contains additivesincluding glycine (pH 2.4) and ascorbic acid to obtain a final pH 7.4and fast reconstitution (~5 min in reconstitution fluid) before use.

Example 5: Freeze-Dried Samples of Biopolymers Mixed With Human Plasmaor Serum

Upon mixing chitosan or hydrophobically-modified (hm) chitosan (each 2wt% solutions in 0.2 M AcOH) with human plasma or human serum, liquidmixtures were obtained. The solutions were poured into weigh boats,which were freeze-dried to remove water. The resulting structures werewafers with dimensions of 50 mm × 50 mm × 4 mm which were beige to ayellowish color, largely dominated by the color of the plasma or serum.The amount of chitosan was about 25 wt% based on the final dry weight ofthe composition, and the amount of the plasma or serum components wasabout 75 wt%.

Mixtures were made in a 50/50 w/w ratio of biopolymer (e.g., chitosan orhm-chitosan) to blood component (plasma or serum); 9.7 g biopolymer to9.7 g of blood component, thus the final weight percent of chitosan inthe solution mixture prior to freeze-drying was about 1%. In controlsamples, only serum or plasma was present, there was no chitosan inthese samples. Samples were freeze-dried for 36 h at -20° C., and thenfor 12 h at + 10° C. at an average vacuum of 100 µbar. Exemplary resultsare shown in Table 2 below.

TABLE 2 Properties of samples of biopolymers mixed with human plasma orserum Sample # Components Color Consistency Integrity 1 Serum onlybeige/yellow Homogeneous with cracks on surface of wafer Easily friable2 Plasma only beige/yellow Homogeneous with cracks on surface of waferEasily friable 3 Serum + Chitosan beige/yellow Homogeneous Moderatelycohesive; cracks with low pressure 4 Plasma + Chitosan beige/yellowHomogeneous Moderately cohesive; cracks with low pressure 5 Serum +hm-Chitosan beige/yellow Homogeneous Cohesive; cracks with forcefulpressure 6 Plasma + hm-chitosan beige/yellow Homogeneous with cracks onsurface of wafer Moderately cohesive; cracks with low pressure

The results indicated that to apply serum as a solid dressing, chitosanor hm-chitosan could be pre-mixed with the serum which thereby created acohesive sponge-like solid. Without the biopolymer, the serum on its ownwas much too friable to apply to an anatomical surface.

Additionally, the chitosan and hm-chitosan formed a less cohesivedressing with plasma than with the serum. This may be related to aninteraction between the chitosan and clotting proteins present in theplasma, but not present in the serum.

Example 6: Synthesis of Hydrophobically-Modified Chitosan

Hydrophobically-modified (hm) chitosan was synthesized by addition ofpalmitic anhydride (0.1 g; TCI America) dissolved in 20 mL EtOH at 70°C. to Chitosan (2.0 g; Chitoclear hqg 400, Primex (Iceland)) dissolvedin 200 mL of a 50/50 v/v mixture of 0.2 M AcOH in water and EtOH at 70°C. The reaction was performed under continuous stirring for 1 hour. Theproduct was precipitated by dropwise addition of 1.0 M NaOH. Finally,the resulting precipitate was washed with 100 mL of EtOH, and driedunder vacuum overnight. The dried pulp was pulverized into powder, whichwas then redissolved in 0.2 M AcOH in water as described earlier in thissection for mixture with blood components.

Example 7: Mixing Serum With Chitosans of Varying Molecular Weights

Upon mixing chitosan (each 2 wt% solutions in 0.2 M AcOH) with humanserum, liquid mixtures were obtained of varying viscosities andmucoadhesive strengths. Chitosans (low molecular weight (~100 kDa),medium molecular weight (~250 kDa) and high molecular weight (~400 kDa))were obtained from Sigma. Zero-shear viscosities were measured with anAR2000 Stress-controlled rheometer under steady-state flow (n=3).Mucoadhesion measurements were performed using a TA-XT Texture Analyzer(Stable Micro Systems) equipped with a load cell of 1 kg (n=3). A/MUCmeasuring system (mucoadhesion test ring) consisting of a ring in whichthe biological support can be fixed and a cylinder probe with a diameterof 1 cm were used. In this study, mucin dispersion (mucin from porcinestomach, Type II, Sigma-Aldrich, Germany) was used instead of abiological support. A filter paper disc was wetted with 10 mL of mucindispersion in phosphate buffer (8%, m/m, pH 6.4). 20 mg of each samplewas applied to the cylinder probe. The sample and biologic substratewere put in contact with a preload of 6000 mN for 3 min. The cylinderprobe was moved upwards at a predetermined speed of 2.5 mm min⁻¹ toseparate the mucoadhesive interface (mucin-sample). Exemplary resultsare shown in Table 3 below.

TABLE 3 Properties of samples of serum mixed with chitosans of varyingmolecular weights Sample # Components Viscosity (mPa•s) Mucoadhesiveness(N mm) 1 Serum only 1.75 ± 0.06 0.001 ± 0.00012 3 Serum + Chitosan (lowmolecular weight) 122 ± 5 0.011 ± 0.001 4 Serum + Chitosan (mediummolecular weight) 276 ± 16 0.021 ± 0.005 5 Serum + Chitosan (highmolecular weight) 1,257 ± 112 0.075 ± 0.007

The data showed a clear increase in viscosity and mucoadhesiveness asthe molecular weight of the chitosan increased. These variables areimportant to the application of the formulation to the eye for optimizedtherapeutic effect via bioavailability of the serum.

As described in detail above, the molecular structure of chitosanenables its properties to be modified. The amount of chitosan added tothe freeze-dried plasma or serum composition can also be adjusted. Theviscosity and the mucoadhesive nature of the freeze-dried plasma orserum composition can be adjusted, i.e., to range from a liquid form(i.e., that could be placed in a bottle as drop), to a film that can bemolded into any shape and be used as a patch and possible even a contactlens. Between the liquid form and the film form there can be made formsthat resemble both gels and ointments.

Depending on the traumatized non-keratinized epithelial surface beingtreated, one form may be more suitable then the other. For example fordry eye a drop may be the most commonly used or preferred form. Forcases of very severe dry eye, a gel and ointment form may be moresuitable. Additionally, a contact lens made out of a film that slowlybiodegrades and releases the serum/plasmas bioactive proteins over timemay be better suited for cases of the most severely traumatized cornealepithelium.

Example 8: Determination of the Glass Transition Temperature (Tg′) andCollapse Temperature of Sucrose Formulation Containing Pooled HumanSerum and Human Plasma Materials Supplied and Preparation

Pooled human serum and pooled human plasma with 4% (w/v) sodium citratewere used. 40 µL (240 mg) of serum or plasma sample was mixed with 20 µLof 15% (w/v) sucrose (0.22 µm filtered) to yield 60 µL of formulation(i.e., 40 mg/mL total protein in 5% (w/v) sucrose).

Methods

Differential Scanning Calorimetry (DSC): A TA Q1000 Modulated DSCequipped with a RCS (refrigerated cooling system) (TA Instruments, NewCastle, DE) was used to determine the glass transition temperature ofthe sucrose formulations containing serum and plasma samples. A 10 µlsample was placed into an aluminum pan and hermetically sealed. The massof the sample, the pan, and the reference pan were used to determinedifferences in heat flow between the sample and reference (an emptypan). Nitrogen (NT 3500) was used as the purge gas with a flow rate of50 mL/min. Samples were run in a conventional DSC mode, with -10° C./min cooling rate from 20° C. to -60° C., isothermal at -60° C. for 30minutes and a scan rate of 2° C./ min during ramp up from -70° C. to +20° C.

Freeze-Drying Microscopy (FDM): Traditional low temperature freezedrying microscopy (FDM) was performed using a temperature and pressurecontrolled FDCS 196 stage (Linkam, Tadworth, Surrey, UK) and an opticalmicroscope (Olympus BH-2, Tokyo, Japan) equipped with a color videocamera (QImaging, Surrey, BC, Canada). A 15 mm round glass cover slidewas placed on the freezing stage block, over a drop of silicon immersionoil which improves the contact and heat transfer from the block to theglass. Then 4.5 uL of sample was pipetted onto the 15 mm glass coverslide as a drop. The sample was subsequently covered with a 9 mm glasscover slide to spread the sample volume evenly between the two glasscover slides. The sample was cooled to -50° C. at 10° C./min andequilibrated for 30 minutes at 200 mTorr vacuum. The sample block wasthen carefully heated to -5° C. at a rate of at 1.0° C./min. Images werecaptured after every 1° C. rise in temperature using a digital camerasystem and were analyzed qualitatively to determine the structuralchanges in the sample. The onset of structural changes indicatescollapse.

Results

The formulation containing serum was mixed thoroughly for 15 secondswith pipet before 10 µL was added to Al pans carefully. At a temperatureof about -35° C., a baseline shift in the heat flow, characteristic ofthe glass transition, was observed and an average glass transitiontemperature of -33.5° C. was obtained during the ramp up (FIG. 2A). Theformulation containing plasma was mixed thoroughly for 15 seconds withpipet before 10 µL was added to Al pans carefully. At a temperature ofabout -35° C., a baseline shift in the heat flow, characteristic of theglass transition, was observed and an average glass transitiontemperature of -33.5° C. was obtained during the ramp up (FIG. 2B).

A formulation of 5% sucrose with serum at -25° C., a collapse front,which indicates a loss of structural integrity, began to appear and thesublimation front eventually collapsed at -23° C. A formulation of 5%sucrose with plasma at -23° C., a collapse front, which indicates a lossof structural integrity, began to appear, and the sublimation fronteventually collapsed at -21° C.

The glass transition temperature (Tg′) for sucrose formulationcontaining serum and plasma were measured using standard DSC and wereobserved to be in between -35° C. to -32° C. range. The structuralchanges in FDM were prominent with a visible sublimation and a classicalcollapse which occurred at -24° C. for the formulation containing serumand -22° C. for the formulation containing plasma.

The disclosure has been described herein by reference to certainembodiments. However, as other variations will become apparent to thoseskilled in the art, the disclosure is not to be considered as limitedthereto. Any compositions or methods provided herein can be combinedwith one or more of any of the other compositions and methods providedherein.

Other features and advantages of the compositions and methods describedherein will be apparent to those skilled in the art from the detaileddescription and claims. All patents, patent applications, and referencescited anywhere are hereby incorporated by reference in their entirety.

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments described herein. Such equivalents are intended to beencompassed by the following claims.

OTHER EMBODIMENTS

It is to be understood that while the disclosure has been described inconjunction with the detailed description thereof, the foregoingdescription is intended to illustrate and not limit the scope of thedisclosure, which is defined by the scope of the appended claims. Otheraspects, advantages, and modifications are within the scope of thefollowing claims.

What is claimed is: 1-70. (canceled)
 71. A pharmaceutical composition comprising serum and a polysaccharide or derivative thereof in an aqueous carrier, wherein the composition comprises up to 90 weight percent (wt%) serum and up to 40.0% polysaccharide, and wherein the polysaccharide is chitosan, cellulose, dextrin, pectin, alginic acid, agar, agarose, carrageenan, or any derivatives thereof.
 72. The pharmaceutical composition of claim 71, wherein the pH of the composition is from 3.5 to 8.0.
 73. The pharmaceutical composition of claim 71, wherein the serum is isolated from blood of a mammal.
 74. The pharmaceutical composition of claim 73, wherein the mammal is a human, sheep, goat, pig, horse, dog, feline, fetal, or cow.
 75. The pharmaceutical composition of claim 74, wherein the serum is human serum.
 76. The pharmaceutical composition of claim 75, wherein the human serum is umbilical cord serum.
 77. The pharmaceutical composition of claim 71, wherein the serum is exposed to one or more of a solvent, a detergent, psoralen, ultraviolet light, filtration, and gamma irradiation, prior to its incorporation into the composition.
 78. The pharmaceutical composition of claim 71, wherein the composition is formulated as a solution, suspension, emulsion, gel, film, patch, cream or ointment.
 79. The pharmaceutical composition of claim 71, further comprising an additive.
 80. The pharmaceutical composition of claim 79, wherein the additive is selected from the group consisting of: chitosan, alginate, gelatin, hyaluronic acid, gellan gum, dextran, polyethylene glycol, polyethylene oxide, glucose, glucosamine, sodium chloride, polylactic acid, polylactic-co-glycolic acid, glycerol, carboxymethylcellulose, hydroxymethylcellulose, glycerin, trehalose, sodium hyaluronate, hydroxypropyl--guar, castor oil, flaxseed oil, and mineral oil.
 81. The pharmaceutical composition of claim 79, wherein the additive comprises one or more of glycine, ascorbic acid, alginate, gelatin, glycol chitosan, lactic acid, acetic acid, hypochlorous acid, citric acid, boric acid, glutamic acid, and glycolic acid.
 82. The pharmaceutical composition of claim 71, wherein the composition is an ophthalmic composition.
 83. A method for treating dry eye syndrome in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the pharmaceutical composition of claim 71, thereby treating dry eye syndrome in the subject.
 84. The method of claim 83, wherein treating dry eye syndrome in the subject comprises treating or ameliorating one or more symptoms associated with risk factors for temporary or chronic dry eye.
 85. The method of claim 84, wherein the risk factors comprise skin disease on or near eyelids, gland disease in eyelids, hormone replacement therapy, refractive surgery, chemical and thermal burns, allergies, infrequent blinking, loss of sensation in cornea, immune system disorder, chronic inflammation in conjunctiva, membrane lining of eyelid, or lacrimal gland, infection, exposure to irritants, and keratitis.
 86. A method for treating an epithelial surface of a subject in need thereof, the method comprising applying a therapeutically effective amount of the pharmaceutical composition of claim 71 to the epithelial surface of the subject, thereby treating the epithelial surface of the subject.
 87. The method of claim 86, wherein the epithelial surface is selected from an ocular surface, an oral surface, a vaginal surface, a skin wound, and the surface of a wound.
 88. The pharmaceutical composition of claim 75, wherein the serum is serum derived from plasma.
 89. The pharmaceutical composition of claim 71, wherein the pharmaceutical composition is a freeze-dried composition. 